chikv Search Results


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  • chikv  (ATCC)
    94
    ATCC chikv
    Viral infection in SPCS1 −/− cells a–c . Alphaviruses replicate and are processed efficiently in 293T cells in the absence of expression of SPCS1. a . SINV infection in control and SPCS1 −/− clonal cells. Cells were infected (MOI of 0.01) and supernatants were harvested and analyzed by FFA. The results are the average of two independent experiments performed in triplicate. b . Control and SPCS1 −/− gene-edited 293T cells were infected with SINV. At the indicated time, lysates were prepared, electrophoresed and Western blotted with anti-SINV E2 ascites fluid (ATCC VR-1248AF). c . Control or SPCS1 −/− 293T cells were infected with <t>CHIKV</t> (MOI of 5). 8 h later, cells were labeled for 30 min with 35 S cysteine-methionine. Excess cold cysteine-methionine was added for indicated chase times (0, 1 or 4 h). An uninfected control established the specificity of the immunoprecipitation. After 35 S labeling, lysates were prepared and immunoprecipitated with anti-E2 MAb (CHK-48). Immunoprecipitates were left untreated (blank) or treated with Endo H (E) or PNGase F (P) for 1 h at 37°C prior to SDS-PAGE and fluorography. d . Sequencing of SPCS1 alleles in gene-edited 293T and Huh7 cell clones after puromycin selection and limiting dilution cloning. The sgRNA targeting site and the “PAM” sequences are highlighted at the top of WT gene, and the sequence of edited alleles are indicated. e . Western blotting of bulk-selected or clonal (clone #7) Huh7.5 cells (control and SPCS1 sgRNA selected) for expression of SPCS1 (~12 kDa). f . WNV, HCV, ZIKV, or <t>JEV</t> (Bennett strain) infection in control and SPCS1-deficient Huh7.5 cells. Cells were infected at an MOI of 0.01 (WNV, ZIKV, JEV) or 1 (HCV) and supernatants were harvested and analyzed by FFA. The results are the average of two independent experiments performed in triplicate. g . Control or SPCS1 −/− Huh7.5 cells were infected at an MOI of 150 for 45 h with a pathogenic JEV isolate (Bennett strain). Lysates were blotted with an anti-JEV E MAb. Higher molecular weight bands (E hi and E med ) that reacted specifically with the anti-E MAb are indicated. One representative experiment of two is shown and loading controls (β-actin) are included. For gel source data, see Supplementary Figure 1 .
    Chikv, supplied by ATCC, used in various techniques. Bioz Stars score: 94/100, based on 114 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    EUROIMMUN chikungunya virus
    Longitudinal variation of the sensitivity of IgM assays in the first seven days after fever onset. 1a: overall sensitivity based on samples collected from first and second <t>chikungunya</t> fever outbreaks in January and May to September 2008, respectively; 1b: sensitivity based on samples collected from the first outbreak due to CHIKV-A226 strain; 1c: sensitivity based on samples collected from the second outbreak due to CHIKV-226V strain. The numbers in brackets along the x-axis represent the number of samples tested for each day after onset of fever.
    Chikungunya Virus, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 89/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Illumina Inc chikv
    <t>CHIKV</t> infection, <t>RNA</t> production and small RNA biogenesis following rearing at 18°C or 28°C. ( A ) Infectivity of CHIKV for Ae. aegypti Liverpool strain following per os challenge. Each bar represents the average of three biological replicates of 27–50 mosquitoes each. Small RNA size distributions of CHIKV vsiRNAs and vpiRNAs ( B ) and absolute quantitation of CHIKV (+) strand RNA ( C ) at 8 hrs post injection into Ae. aegypti Liverpool strain females reared at 18°C or 28°C. Error bars indicate the standard deviation amongst three biological replicates. Significance was assessed via a two-tailed Student's t-test.
    Chikv, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 92/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Abcam eilv chikv
    Comparison of <t>EILV/CHIKV</t> and Abcam human anti-CHIKV IgM-capture ELISA methods. (A) raw OD values or (B) signal-to-noise ratios (calculated by dividing positive sample OD values by mean negative control OD value) produced by samples of varying degrees of seropositivity were compared. Low-positive samples had PRNT 50 ≤80 (n = 4), medium-positive samples had PRNT 50 160–320 (n = 4), and high-positive samples had PRNT 50 > 640 (n = 24). Mean and standard deviations are reported. A 2-way ANOVA with Tukey’s test was used to compare absorbance values and signal-to-noise ratios within each ELISA method. 2-way ANOVA with Bonferroni’s test was used to compare absorbance values and signal-to-noise ratios between each ELISA method.
    Eilv Chikv, supplied by Abcam, used in various techniques. Bioz Stars score: 88/100, based on 36 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    EUROIMMUN anti chikv elisa igm
    Comparison of sensitivity by serum titration. Chikungunya virus <t>(CHIKV)</t> <t>IgM+</t> samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.
    Anti Chikv Elisa Igm, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 90/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam anti chikv igm human elisa kit
    Comparison of sensitivity by serum titration. Chikungunya virus <t>(CHIKV)</t> <t>IgM+</t> samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.
    Anti Chikv Igm Human Elisa Kit, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    EUROIMMUN chikv igm
    Comparison of sensitivity by serum titration. Chikungunya virus <t>(CHIKV)</t> <t>IgM+</t> samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.
    Chikv Igm, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 90/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    EUROIMMUN anti chikungunya virus elisa igm kit
    Dengue virus (DENV) serological assays. (A) Amount of <t>IgM</t> reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (B) Amount of IgM reactive to DENV in the acute and convalescent samples of participants infected with <t>chikungunya</t> virus (CHIKV). (C) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (D) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with chikungunya virus (CHIKV). <t>ELISA</t> optical density was converted in Panbio units. Asterisks reflect the level of significance between groups after paired T -test was performed: ** p
    Anti Chikungunya Virus Elisa Igm Kit, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    EUROIMMUN anti chikv elisa
    Dengue virus (DENV) serological assays. (A) Amount of <t>IgM</t> reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (B) Amount of IgM reactive to DENV in the acute and convalescent samples of participants infected with <t>chikungunya</t> virus (CHIKV). (C) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (D) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with chikungunya virus (CHIKV). <t>ELISA</t> optical density was converted in Panbio units. Asterisks reflect the level of significance between groups after paired T -test was performed: ** p
    Anti Chikv Elisa, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    EUROIMMUN anti chikv igm antibodies
    Numbers of <t>CHIKV</t> cases, anti-CHIKV <t>IgM</t> status and monthly precipitation in the Chapada district between October 2014 and November 2015.
    Anti Chikv Igm Antibodies, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 93/100, based on 26 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    chikv  (Abcam)
    90
    Abcam chikv
    Numbers of <t>CHIKV</t> cases, anti-CHIKV <t>IgM</t> status and monthly precipitation in the Chapada district between October 2014 and November 2015.
    Chikv, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Focus Diagnostics Inc simplexa chikv kit
    Numbers of <t>CHIKV</t> cases, anti-CHIKV <t>IgM</t> status and monthly precipitation in the Chapada district between October 2014 and November 2015.
    Simplexa Chikv Kit, supplied by Focus Diagnostics Inc, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti chikv mouse monoclonal antibody
    Numbers of <t>CHIKV</t> cases, anti-CHIKV <t>IgM</t> status and monthly precipitation in the Chapada district between October 2014 and November 2015.
    Anti Chikv Mouse Monoclonal Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    ATCC chikv strain s27
    Numbers of <t>CHIKV</t> cases, anti-CHIKV <t>IgM</t> status and monthly precipitation in the Chapada district between October 2014 and November 2015.
    Chikv Strain S27, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    BEI Resources chikv
    Numbers of <t>CHIKV</t> cases, anti-CHIKV <t>IgM</t> status and monthly precipitation in the Chapada district between October 2014 and November 2015.
    Chikv, supplied by BEI Resources, used in various techniques. Bioz Stars score: 94/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Focus Diagnostics Inc chikv
    Analysis of the sensitivity and specificity of the BOB NS1 ELISA. ( A ) The values of BOB inhibition in samples from subjects diagnosed with ZIKV or <t>DENV</t> infection by RT-PCR are shown. The DENV group consists of samples from primary and secondary DENV cases from Nicaragua and Italy. The control group identified as “all” comprises DENV samples as well as samples from patients infected (or vaccinated) with other flaviviruses (i.e., WNV and YFV) or with <t>CHIKV,</t> samples from patients with other illnesses and samples from healthy blood donors. ( B ) Sensitivity calculated on the basis of different time-point cutoff values, and specificity calculated using different control groups. The flaviviruses group comprises samples from DENV and WNV cases and YFV vaccinees. The control group identified as “+ other illnesses” comprises the samples from the flavivirus group, as well as samples with CHIKV and other illnesses (Brazilian samples collected before 2014).
    Chikv, supplied by Focus Diagnostics Inc, used in various techniques. Bioz Stars score: 91/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam human anti chikungunya virus igg elisa kit
    Diversity of the antibody response patterns in <t>chikungunya</t> acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and <t>IgG</t> (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).
    Human Anti Chikungunya Virus Igg Elisa Kit, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    IBT Bioservices rabbit polyclonal anti chikv 181 25
    Diversity of the antibody response patterns in <t>chikungunya</t> acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and <t>IgG</t> (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).
    Rabbit Polyclonal Anti Chikv 181 25, supplied by IBT Bioservices, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Abcam anti chikv igg
    Diversity of the antibody response patterns in <t>chikungunya</t> acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and <t>IgG</t> (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).
    Anti Chikv Igg, supplied by Abcam, used in various techniques. Bioz Stars score: 93/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    EUROIMMUN anti chikv indirect immunofluorescence assay
    Diversity of the antibody response patterns in <t>chikungunya</t> acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and <t>IgG</t> (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).
    Anti Chikv Indirect Immunofluorescence Assay, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 86/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    EUROIMMUN anti chikv iift kit
    Diversity of the antibody response patterns in <t>chikungunya</t> acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and <t>IgG</t> (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).
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    92
    EUROIMMUN anti chikungunya virus immunofluorescent assay ifa
    Diversity of the antibody response patterns in <t>chikungunya</t> acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and <t>IgG</t> (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).
    Anti Chikungunya Virus Immunofluorescent Assay Ifa, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 92/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Viral infection in SPCS1 −/− cells a–c . Alphaviruses replicate and are processed efficiently in 293T cells in the absence of expression of SPCS1. a . SINV infection in control and SPCS1 −/− clonal cells. Cells were infected (MOI of 0.01) and supernatants were harvested and analyzed by FFA. The results are the average of two independent experiments performed in triplicate. b . Control and SPCS1 −/− gene-edited 293T cells were infected with SINV. At the indicated time, lysates were prepared, electrophoresed and Western blotted with anti-SINV E2 ascites fluid (ATCC VR-1248AF). c . Control or SPCS1 −/− 293T cells were infected with CHIKV (MOI of 5). 8 h later, cells were labeled for 30 min with 35 S cysteine-methionine. Excess cold cysteine-methionine was added for indicated chase times (0, 1 or 4 h). An uninfected control established the specificity of the immunoprecipitation. After 35 S labeling, lysates were prepared and immunoprecipitated with anti-E2 MAb (CHK-48). Immunoprecipitates were left untreated (blank) or treated with Endo H (E) or PNGase F (P) for 1 h at 37°C prior to SDS-PAGE and fluorography. d . Sequencing of SPCS1 alleles in gene-edited 293T and Huh7 cell clones after puromycin selection and limiting dilution cloning. The sgRNA targeting site and the “PAM” sequences are highlighted at the top of WT gene, and the sequence of edited alleles are indicated. e . Western blotting of bulk-selected or clonal (clone #7) Huh7.5 cells (control and SPCS1 sgRNA selected) for expression of SPCS1 (~12 kDa). f . WNV, HCV, ZIKV, or JEV (Bennett strain) infection in control and SPCS1-deficient Huh7.5 cells. Cells were infected at an MOI of 0.01 (WNV, ZIKV, JEV) or 1 (HCV) and supernatants were harvested and analyzed by FFA. The results are the average of two independent experiments performed in triplicate. g . Control or SPCS1 −/− Huh7.5 cells were infected at an MOI of 150 for 45 h with a pathogenic JEV isolate (Bennett strain). Lysates were blotted with an anti-JEV E MAb. Higher molecular weight bands (E hi and E med ) that reacted specifically with the anti-E MAb are indicated. One representative experiment of two is shown and loading controls (β-actin) are included. For gel source data, see Supplementary Figure 1 .

    Journal: Nature

    Article Title: A CRISPR screen defines a signal peptide processing pathway required by flaviviruses

    doi: 10.1038/nature18625

    Figure Lengend Snippet: Viral infection in SPCS1 −/− cells a–c . Alphaviruses replicate and are processed efficiently in 293T cells in the absence of expression of SPCS1. a . SINV infection in control and SPCS1 −/− clonal cells. Cells were infected (MOI of 0.01) and supernatants were harvested and analyzed by FFA. The results are the average of two independent experiments performed in triplicate. b . Control and SPCS1 −/− gene-edited 293T cells were infected with SINV. At the indicated time, lysates were prepared, electrophoresed and Western blotted with anti-SINV E2 ascites fluid (ATCC VR-1248AF). c . Control or SPCS1 −/− 293T cells were infected with CHIKV (MOI of 5). 8 h later, cells were labeled for 30 min with 35 S cysteine-methionine. Excess cold cysteine-methionine was added for indicated chase times (0, 1 or 4 h). An uninfected control established the specificity of the immunoprecipitation. After 35 S labeling, lysates were prepared and immunoprecipitated with anti-E2 MAb (CHK-48). Immunoprecipitates were left untreated (blank) or treated with Endo H (E) or PNGase F (P) for 1 h at 37°C prior to SDS-PAGE and fluorography. d . Sequencing of SPCS1 alleles in gene-edited 293T and Huh7 cell clones after puromycin selection and limiting dilution cloning. The sgRNA targeting site and the “PAM” sequences are highlighted at the top of WT gene, and the sequence of edited alleles are indicated. e . Western blotting of bulk-selected or clonal (clone #7) Huh7.5 cells (control and SPCS1 sgRNA selected) for expression of SPCS1 (~12 kDa). f . WNV, HCV, ZIKV, or JEV (Bennett strain) infection in control and SPCS1-deficient Huh7.5 cells. Cells were infected at an MOI of 0.01 (WNV, ZIKV, JEV) or 1 (HCV) and supernatants were harvested and analyzed by FFA. The results are the average of two independent experiments performed in triplicate. g . Control or SPCS1 −/− Huh7.5 cells were infected at an MOI of 150 for 45 h with a pathogenic JEV isolate (Bennett strain). Lysates were blotted with an anti-JEV E MAb. Higher molecular weight bands (E hi and E med ) that reacted specifically with the anti-E MAb are indicated. One representative experiment of two is shown and loading controls (β-actin) are included. For gel source data, see Supplementary Figure 1 .

    Article Snippet: Cells then were rinsed one additional time with Perm buffer, transferred to a U-bottom plate, and incubated for 1 h at 4°C with 1 µg/ml of the following virus-specific antibodies: WNV (human E16 ); DENV2 (mouse E18 ); JEV (mouse E18 ); YFV (mouse E60 ); CHIKV (CHK-11 ); SINV (ascites fluid, ATCC VR-1248AF), LACV (807-31 and 807-33, gift of A. Pekosz, Baltimore, MD).

    Techniques: Infection, Expressing, Western Blot, Labeling, Immunoprecipitation, SDS Page, Sequencing, Clone Assay, Selection, Molecular Weight

    SPCS1 is required for flavivirus protein processing and infection a . Western blotting of SPCS1 −/− 293T cells. b . Cells were transfected with YFV-luciferase replicon RNA (WT GDD or loss-of-function GVD). Firefly luciferase activity was measured and normalized to intracellular protein levels. The data reflects the average of two to three independent experiments performed in duplicate. c–h . Cells were infected with WNV ( c, h ), DENV-2 ( d ), JEV ( e ), YFV ( f ) or ZIKV ( g ), and viral yield measured. In h , cells were trans-complemented with an SPCS1 or control plasmid. Results are the average of two to three independent experiments performed in triplicate. i–k . Cells were infected with CHIKV (alphavirus), RVFV (bunyavirus), or VSV (rhabdovirus) and viral yield was measured. Results are the average of two to three independent experiments performed in triplicate. l . The polyprotein processing strategy of flaviviruses 13 . Red and blue arrows indicate sites of cleavage by host and viral (NS2B-NS3) proteases, respectively. m–o . Control or SPCS1 −/− 293T ( m , o ) or Huh7.5 ( n ) cells were infected with WNV ( m, o ) or JEV ( n ). Lysates were blotted with ( m ) anti-WNV E, ( n ) anti-JEV E, or ( o ) anti-WNV NS1 MAbs. Higher molecular weight bands (E hi , E med , and NS1 hi ) that react with anti-flavivirus MAbs are indicated. One experiment of three is shown. p . 293T cells were infected with CHIKV or RVFV. Lysates were blotted with anti-CHIKV E2 or anti-RVFV Gn mAbs. One experiment of two is shown. For gel source data, see Supplementary Figure 1 .

    Journal: Nature

    Article Title: A CRISPR screen defines a signal peptide processing pathway required by flaviviruses

    doi: 10.1038/nature18625

    Figure Lengend Snippet: SPCS1 is required for flavivirus protein processing and infection a . Western blotting of SPCS1 −/− 293T cells. b . Cells were transfected with YFV-luciferase replicon RNA (WT GDD or loss-of-function GVD). Firefly luciferase activity was measured and normalized to intracellular protein levels. The data reflects the average of two to three independent experiments performed in duplicate. c–h . Cells were infected with WNV ( c, h ), DENV-2 ( d ), JEV ( e ), YFV ( f ) or ZIKV ( g ), and viral yield measured. In h , cells were trans-complemented with an SPCS1 or control plasmid. Results are the average of two to three independent experiments performed in triplicate. i–k . Cells were infected with CHIKV (alphavirus), RVFV (bunyavirus), or VSV (rhabdovirus) and viral yield was measured. Results are the average of two to three independent experiments performed in triplicate. l . The polyprotein processing strategy of flaviviruses 13 . Red and blue arrows indicate sites of cleavage by host and viral (NS2B-NS3) proteases, respectively. m–o . Control or SPCS1 −/− 293T ( m , o ) or Huh7.5 ( n ) cells were infected with WNV ( m, o ) or JEV ( n ). Lysates were blotted with ( m ) anti-WNV E, ( n ) anti-JEV E, or ( o ) anti-WNV NS1 MAbs. Higher molecular weight bands (E hi , E med , and NS1 hi ) that react with anti-flavivirus MAbs are indicated. One experiment of three is shown. p . 293T cells were infected with CHIKV or RVFV. Lysates were blotted with anti-CHIKV E2 or anti-RVFV Gn mAbs. One experiment of two is shown. For gel source data, see Supplementary Figure 1 .

    Article Snippet: Cells then were rinsed one additional time with Perm buffer, transferred to a U-bottom plate, and incubated for 1 h at 4°C with 1 µg/ml of the following virus-specific antibodies: WNV (human E16 ); DENV2 (mouse E18 ); JEV (mouse E18 ); YFV (mouse E60 ); CHIKV (CHK-11 ); SINV (ascites fluid, ATCC VR-1248AF), LACV (807-31 and 807-33, gift of A. Pekosz, Baltimore, MD).

    Techniques: Infection, Western Blot, Transfection, Luciferase, Activity Assay, Plasmid Preparation, Molecular Weight

    Gene editing of SEC11A and SEC11C do not susbtantively impact infection of several enveloped viruses ( Top ) 293T cells were administered the indicated sgRNA and isolated in bulk after puromycin drug selection. Western blotting confirmed gene editing of SEC11A ( left , 20 kDa) or SEC11C ( middle , 22 kDa). No difference in levels or migration pattern of WNV E was observed in SEC11A or SEC11C gene edited cells ( right ) after WNV infection at an MOI of 200 for 24 h. Spaces between the Western blots indicate cropping to remove lanes that were not relevant to this experiment. ( Bottom ) Control or gene-edited 293T cells were infected with viruses and supernatants were harvested after infection for titration. Left . WNV (MOI of 0.01, 72 h) or YFV (MOI of 1, 72 h); Right , SINV (MOI of 0.01, 72 h), CHIKV (MOI of 0.01, 36 h), VSV (MOI of 0.01, 36 h), or RVFV (MOI of 1, 72 h). Results are representative of two independent experiments. For gel source data, see Supplementary Figure 1 .

    Journal: Nature

    Article Title: A CRISPR screen defines a signal peptide processing pathway required by flaviviruses

    doi: 10.1038/nature18625

    Figure Lengend Snippet: Gene editing of SEC11A and SEC11C do not susbtantively impact infection of several enveloped viruses ( Top ) 293T cells were administered the indicated sgRNA and isolated in bulk after puromycin drug selection. Western blotting confirmed gene editing of SEC11A ( left , 20 kDa) or SEC11C ( middle , 22 kDa). No difference in levels or migration pattern of WNV E was observed in SEC11A or SEC11C gene edited cells ( right ) after WNV infection at an MOI of 200 for 24 h. Spaces between the Western blots indicate cropping to remove lanes that were not relevant to this experiment. ( Bottom ) Control or gene-edited 293T cells were infected with viruses and supernatants were harvested after infection for titration. Left . WNV (MOI of 0.01, 72 h) or YFV (MOI of 1, 72 h); Right , SINV (MOI of 0.01, 72 h), CHIKV (MOI of 0.01, 36 h), VSV (MOI of 0.01, 36 h), or RVFV (MOI of 1, 72 h). Results are representative of two independent experiments. For gel source data, see Supplementary Figure 1 .

    Article Snippet: Cells then were rinsed one additional time with Perm buffer, transferred to a U-bottom plate, and incubated for 1 h at 4°C with 1 µg/ml of the following virus-specific antibodies: WNV (human E16 ); DENV2 (mouse E18 ); JEV (mouse E18 ); YFV (mouse E60 ); CHIKV (CHK-11 ); SINV (ascites fluid, ATCC VR-1248AF), LACV (807-31 and 807-33, gift of A. Pekosz, Baltimore, MD).

    Techniques: Infection, Isolation, Selection, Western Blot, Migration, Titration

    Longitudinal variation of the sensitivity of IgM assays in the first seven days after fever onset. 1a: overall sensitivity based on samples collected from first and second chikungunya fever outbreaks in January and May to September 2008, respectively; 1b: sensitivity based on samples collected from the first outbreak due to CHIKV-A226 strain; 1c: sensitivity based on samples collected from the second outbreak due to CHIKV-226V strain. The numbers in brackets along the x-axis represent the number of samples tested for each day after onset of fever.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Evaluation of Chikungunya Diagnostic Assays: Differences in Sensitivity of Serology Assays in Two Independent Outbreaks

    doi: 10.1371/journal.pntd.0000753

    Figure Lengend Snippet: Longitudinal variation of the sensitivity of IgM assays in the first seven days after fever onset. 1a: overall sensitivity based on samples collected from first and second chikungunya fever outbreaks in January and May to September 2008, respectively; 1b: sensitivity based on samples collected from the first outbreak due to CHIKV-A226 strain; 1c: sensitivity based on samples collected from the second outbreak due to CHIKV-226V strain. The numbers in brackets along the x-axis represent the number of samples tested for each day after onset of fever.

    Article Snippet: Using sera collected in outbreaks caused by two variants of Chikungunya virus (A226 and 226V), we tested 2 commercial IgM tests (CTK lateral flow rapid test and EUROIMMUN IFA) alongside our in-house IgM assays (using both variants of the virus).

    Techniques:

    CHIKV infection, RNA production and small RNA biogenesis following rearing at 18°C or 28°C. ( A ) Infectivity of CHIKV for Ae. aegypti Liverpool strain following per os challenge. Each bar represents the average of three biological replicates of 27–50 mosquitoes each. Small RNA size distributions of CHIKV vsiRNAs and vpiRNAs ( B ) and absolute quantitation of CHIKV (+) strand RNA ( C ) at 8 hrs post injection into Ae. aegypti Liverpool strain females reared at 18°C or 28°C. Error bars indicate the standard deviation amongst three biological replicates. Significance was assessed via a two-tailed Student's t-test.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Cooler Temperatures Destabilize RNA Interference and Increase Susceptibility of Disease Vector Mosquitoes to Viral Infection

    doi: 10.1371/journal.pntd.0002239

    Figure Lengend Snippet: CHIKV infection, RNA production and small RNA biogenesis following rearing at 18°C or 28°C. ( A ) Infectivity of CHIKV for Ae. aegypti Liverpool strain following per os challenge. Each bar represents the average of three biological replicates of 27–50 mosquitoes each. Small RNA size distributions of CHIKV vsiRNAs and vpiRNAs ( B ) and absolute quantitation of CHIKV (+) strand RNA ( C ) at 8 hrs post injection into Ae. aegypti Liverpool strain females reared at 18°C or 28°C. Error bars indicate the standard deviation amongst three biological replicates. Significance was assessed via a two-tailed Student's t-test.

    Article Snippet: Small RNA libraries derived from whole female Ae. aegypti infected with CHIKV (three biological replicates per temperature) were barcoded and constructed with Illumina's TruSeq™ small RNA prep kit and sequenced on an Illumina HiSeq.

    Techniques: Infection, Quantitation Assay, Injection, Standard Deviation, Two Tailed Test

    Comparison of EILV/CHIKV and Abcam human anti-CHIKV IgM-capture ELISA methods. (A) raw OD values or (B) signal-to-noise ratios (calculated by dividing positive sample OD values by mean negative control OD value) produced by samples of varying degrees of seropositivity were compared. Low-positive samples had PRNT 50 ≤80 (n = 4), medium-positive samples had PRNT 50 160–320 (n = 4), and high-positive samples had PRNT 50 > 640 (n = 24). Mean and standard deviations are reported. A 2-way ANOVA with Tukey’s test was used to compare absorbance values and signal-to-noise ratios within each ELISA method. 2-way ANOVA with Bonferroni’s test was used to compare absorbance values and signal-to-noise ratios between each ELISA method.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Comparison of EILV/CHIKV and Abcam human anti-CHIKV IgM-capture ELISA methods. (A) raw OD values or (B) signal-to-noise ratios (calculated by dividing positive sample OD values by mean negative control OD value) produced by samples of varying degrees of seropositivity were compared. Low-positive samples had PRNT 50 ≤80 (n = 4), medium-positive samples had PRNT 50 160–320 (n = 4), and high-positive samples had PRNT 50 > 640 (n = 24). Mean and standard deviations are reported. A 2-way ANOVA with Tukey’s test was used to compare absorbance values and signal-to-noise ratios within each ELISA method. 2-way ANOVA with Bonferroni’s test was used to compare absorbance values and signal-to-noise ratios between each ELISA method.

    Article Snippet: Plates were coated with EILV/CHIKV at 5 x 104 PFU per well and antigen was detected by both polyclonal and monoclonal antibodies with dilutions from 1:50 to 1:51,200.

    Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Produced, Plaque Reduction Neutralization Test

    Indirect mouse anti-CHIKV IgG ELISAs utilizing either (A) EILV/CHIKV or (B) cell-lysate antigen to detect serially diluted polyclonal anti-CHIKV MIAF (measured over a range of serum dilutions, red) or monoclonal antibody CHK-175 (expressed in ng quantities, blue). MIAF against Gamboa virus and MIS against mosquito antigens were included as negative controls. Mean and standard deviation of 2 replicates are reported.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Indirect mouse anti-CHIKV IgG ELISAs utilizing either (A) EILV/CHIKV or (B) cell-lysate antigen to detect serially diluted polyclonal anti-CHIKV MIAF (measured over a range of serum dilutions, red) or monoclonal antibody CHK-175 (expressed in ng quantities, blue). MIAF against Gamboa virus and MIS against mosquito antigens were included as negative controls. Mean and standard deviation of 2 replicates are reported.

    Article Snippet: Plates were coated with EILV/CHIKV at 5 x 104 PFU per well and antigen was detected by both polyclonal and monoclonal antibodies with dilutions from 1:50 to 1:51,200.

    Techniques: Standard Deviation

    Stability of EILV/CHIKV in different buffers determined by accelerated decay at elevated temperature. Mean and standard deviations of three replicates are reported. A 2-way ANOVA with Tukey’s test was used to compare the effects of different buffer preparations on ELISA activity.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Stability of EILV/CHIKV in different buffers determined by accelerated decay at elevated temperature. Mean and standard deviations of three replicates are reported. A 2-way ANOVA with Tukey’s test was used to compare the effects of different buffer preparations on ELISA activity.

    Article Snippet: Plates were coated with EILV/CHIKV at 5 x 104 PFU per well and antigen was detected by both polyclonal and monoclonal antibodies with dilutions from 1:50 to 1:51,200.

    Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay

    Comparison of EILV/CHIKV and Abcam human anti-CHIKV IgG-capture ELISA methods. (A) raw OD values or (B) signal-to-noise ratios (calculated by dividing positive sample OD values by mean negative control OD value) produced by samples of varying degrees of seropositivity were compared. Low-positive samples had PRNT 80 ≤80 (n = 10), medium-positive samples had PRNT 80 160–320 (n = 10), and high-positive samples had PRNT 80 > 640 (n = 10). Means and standard deviations are reported. A 2-way ANOVA with Tukey’s test was used to compare absorbance values and signal-to-noise ratios within each ELISA method. 2-way ANOVA with Bonferroni’s test was used to compare absorbance values and signal-to-noise ratios between each ELISA method.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Comparison of EILV/CHIKV and Abcam human anti-CHIKV IgG-capture ELISA methods. (A) raw OD values or (B) signal-to-noise ratios (calculated by dividing positive sample OD values by mean negative control OD value) produced by samples of varying degrees of seropositivity were compared. Low-positive samples had PRNT 80 ≤80 (n = 10), medium-positive samples had PRNT 80 160–320 (n = 10), and high-positive samples had PRNT 80 > 640 (n = 10). Means and standard deviations are reported. A 2-way ANOVA with Tukey’s test was used to compare absorbance values and signal-to-noise ratios within each ELISA method. 2-way ANOVA with Bonferroni’s test was used to compare absorbance values and signal-to-noise ratios between each ELISA method.

    Article Snippet: Plates were coated with EILV/CHIKV at 5 x 104 PFU per well and antigen was detected by both polyclonal and monoclonal antibodies with dilutions from 1:50 to 1:51,200.

    Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Produced, Plaque Reduction Neutralization Test

    Safety of EILV/CHIKV in the newborn mouse model of neurovirulence. (A) Survival of infant mice infected IC with EILV/CHIKV, live-attenuated vaccine strain 181/25, or PBS. (B) Replication of EILV/CHIKV or strain 181/25 in newborn mouse brain tissue.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Safety of EILV/CHIKV in the newborn mouse model of neurovirulence. (A) Survival of infant mice infected IC with EILV/CHIKV, live-attenuated vaccine strain 181/25, or PBS. (B) Replication of EILV/CHIKV or strain 181/25 in newborn mouse brain tissue.

    Article Snippet: Plates were coated with EILV/CHIKV at 5 x 104 PFU per well and antigen was detected by both polyclonal and monoclonal antibodies with dilutions from 1:50 to 1:51,200.

    Techniques: Mouse Assay, Infection

    Human anti-CHIKV antibody-capture ELISAs utilizing EILV/CHIKV as antigen to detect either (A) IgM or (B) IgG antibodies (both measured over a range of serum dilutions, red). Human serum samples that were antibody-positive for either DENV or VEEV but negative for CHIKV by HI were included as negative controls. Mean and standard deviations of 2 replicates are reported.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Human anti-CHIKV antibody-capture ELISAs utilizing EILV/CHIKV as antigen to detect either (A) IgM or (B) IgG antibodies (both measured over a range of serum dilutions, red). Human serum samples that were antibody-positive for either DENV or VEEV but negative for CHIKV by HI were included as negative controls. Mean and standard deviations of 2 replicates are reported.

    Article Snippet: Plates were coated with EILV/CHIKV at 5 x 104 PFU per well and antigen was detected by both polyclonal and monoclonal antibodies with dilutions from 1:50 to 1:51,200.

    Techniques:

    Comparison of sensitivity by serum titration. Chikungunya virus (CHIKV) IgM+ samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Comparison of sensitivity by serum titration. Chikungunya virus (CHIKV) IgM+ samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.

    Article Snippet: In summary, the Euroimmun Anti-CHIKV ELISA (IgM), InBios CHIKjj Detect MAC-ELISA, Abcam Anti-CHIKV IgM human ELISA (with CHIKV-specific IgM serum controls), and Euroimmun Anti-CHIKV IIFT kits were shown to have equivalent performance to the reference assays by which they were evaluated.

    Techniques: Titration

    Performance of CHIKV IgM detection assays compared with CDC results as the reference standard. *CDC CHIKV IHPC had negative result in test. **CDC CHIKV IHPC had positive result in test. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control; ND = not done; RT = rapid test.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Performance of CHIKV IgM detection assays compared with CDC results as the reference standard. *CDC CHIKV IHPC had negative result in test. **CDC CHIKV IHPC had positive result in test. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control; ND = not done; RT = rapid test.

    Article Snippet: In summary, the Euroimmun Anti-CHIKV ELISA (IgM), InBios CHIKjj Detect MAC-ELISA, Abcam Anti-CHIKV IgM human ELISA (with CHIKV-specific IgM serum controls), and Euroimmun Anti-CHIKV IIFT kits were shown to have equivalent performance to the reference assays by which they were evaluated.

    Techniques: Positive Control

    Abcam CHIK MAC-ELISA performance assessment with CDC CHIKV IHPC, by lot number. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Abcam CHIK MAC-ELISA performance assessment with CDC CHIKV IHPC, by lot number. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control.

    Article Snippet: In summary, the Euroimmun Anti-CHIKV ELISA (IgM), InBios CHIKjj Detect MAC-ELISA, Abcam Anti-CHIKV IgM human ELISA (with CHIKV-specific IgM serum controls), and Euroimmun Anti-CHIKV IIFT kits were shown to have equivalent performance to the reference assays by which they were evaluated.

    Techniques: Enzyme-linked Immunosorbent Assay, Positive Control

    Comparison of sensitivity by serum titration. Chikungunya virus (CHIKV) IgM+ samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Comparison of sensitivity by serum titration. Chikungunya virus (CHIKV) IgM+ samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.

    Article Snippet: The Abcam Anti-CHIKV IgM human ELISA kit was evaluated at CDC and CARPHA.

    Techniques: Titration

    Performance of CHIKV IgM detection assays compared with CDC results as the reference standard. *CDC CHIKV IHPC had negative result in test. **CDC CHIKV IHPC had positive result in test. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control; ND = not done; RT = rapid test.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Performance of CHIKV IgM detection assays compared with CDC results as the reference standard. *CDC CHIKV IHPC had negative result in test. **CDC CHIKV IHPC had positive result in test. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control; ND = not done; RT = rapid test.

    Article Snippet: The Abcam Anti-CHIKV IgM human ELISA kit was evaluated at CDC and CARPHA.

    Techniques: Positive Control

    Abcam CHIK MAC-ELISA performance assessment with CDC CHIKV IHPC, by lot number. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Abcam CHIK MAC-ELISA performance assessment with CDC CHIKV IHPC, by lot number. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control.

    Article Snippet: The Abcam Anti-CHIKV IgM human ELISA kit was evaluated at CDC and CARPHA.

    Techniques: Enzyme-linked Immunosorbent Assay, Positive Control

    Comparison of EILV/CHIKV and Abcam human anti-CHIKV IgM-capture ELISA methods. (A) raw OD values or (B) signal-to-noise ratios (calculated by dividing positive sample OD values by mean negative control OD value) produced by samples of varying degrees of seropositivity were compared. Low-positive samples had PRNT 50 ≤80 (n = 4), medium-positive samples had PRNT 50 160–320 (n = 4), and high-positive samples had PRNT 50 > 640 (n = 24). Mean and standard deviations are reported. A 2-way ANOVA with Tukey’s test was used to compare absorbance values and signal-to-noise ratios within each ELISA method. 2-way ANOVA with Bonferroni’s test was used to compare absorbance values and signal-to-noise ratios between each ELISA method.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Comparison of EILV/CHIKV and Abcam human anti-CHIKV IgM-capture ELISA methods. (A) raw OD values or (B) signal-to-noise ratios (calculated by dividing positive sample OD values by mean negative control OD value) produced by samples of varying degrees of seropositivity were compared. Low-positive samples had PRNT 50 ≤80 (n = 4), medium-positive samples had PRNT 50 160–320 (n = 4), and high-positive samples had PRNT 50 > 640 (n = 24). Mean and standard deviations are reported. A 2-way ANOVA with Tukey’s test was used to compare absorbance values and signal-to-noise ratios within each ELISA method. 2-way ANOVA with Bonferroni’s test was used to compare absorbance values and signal-to-noise ratios between each ELISA method.

    Article Snippet: Commercially available anti-CHIKV IgM (ab177848, Lot: GR195090-3, Abcam, Cambridge, MA) and anti-CHIKV IgG Human ELISA Kits (ab177835, Lot: GR148047-1, Abcam) were used according to manufacturer’s instructions.

    Techniques: Enzyme-linked Immunosorbent Assay, Negative Control, Produced, Plaque Reduction Neutralization Test

    Human anti-CHIKV antibody-capture ELISAs utilizing EILV/CHIKV as antigen to detect either (A) IgM or (B) IgG antibodies (both measured over a range of serum dilutions, red). Human serum samples that were antibody-positive for either DENV or VEEV but negative for CHIKV by HI were included as negative controls. Mean and standard deviations of 2 replicates are reported.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Utilization of an Eilat Virus-Based Chimera for Serological Detection of Chikungunya Infection

    doi: 10.1371/journal.pntd.0004119

    Figure Lengend Snippet: Human anti-CHIKV antibody-capture ELISAs utilizing EILV/CHIKV as antigen to detect either (A) IgM or (B) IgG antibodies (both measured over a range of serum dilutions, red). Human serum samples that were antibody-positive for either DENV or VEEV but negative for CHIKV by HI were included as negative controls. Mean and standard deviations of 2 replicates are reported.

    Article Snippet: Commercially available anti-CHIKV IgM (ab177848, Lot: GR195090-3, Abcam, Cambridge, MA) and anti-CHIKV IgG Human ELISA Kits (ab177835, Lot: GR148047-1, Abcam) were used according to manufacturer’s instructions.

    Techniques:

    Comparison of sensitivity by serum titration. Chikungunya virus (CHIKV) IgM+ samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Comparison of sensitivity by serum titration. Chikungunya virus (CHIKV) IgM+ samples from the Caribbean were diluted 2-fold to 1:12,800 in sample dilution buffer. The revised Abcam kit with CHIKV-specific IgM serum controls was used for the evaluation.

    Article Snippet: To compare performance between the kits, the results of 68 samples (35 CDC CHIKV IgM+, 33 CHIKV IgM−) that had been tested in the Euroimmun, InBios, and revised Abcam ELISAs were used ( ).

    Techniques: Titration

    Performance of CHIKV IgM detection assays compared with CDC results as the reference standard. *CDC CHIKV IHPC had negative result in test. **CDC CHIKV IHPC had positive result in test. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control; ND = not done; RT = rapid test.

    Journal: The American Journal of Tropical Medicine and Hygiene

    Article Title: Evaluation of Commercially Available Chikungunya Virus Immunoglobulin M Detection Assays

    doi: 10.4269/ajtmh.16-0013

    Figure Lengend Snippet: Performance of CHIKV IgM detection assays compared with CDC results as the reference standard. *CDC CHIKV IHPC had negative result in test. **CDC CHIKV IHPC had positive result in test. CDC = Centers for Disease Control and Prevention; CHIKV = chikungunya virus; IHPC = in-house positive control; ND = not done; RT = rapid test.

    Article Snippet: To compare performance between the kits, the results of 68 samples (35 CDC CHIKV IgM+, 33 CHIKV IgM−) that had been tested in the Euroimmun, InBios, and revised Abcam ELISAs were used ( ).

    Techniques: Positive Control

    Dengue virus (DENV) serological assays. (A) Amount of IgM reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (B) Amount of IgM reactive to DENV in the acute and convalescent samples of participants infected with chikungunya virus (CHIKV). (C) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (D) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with chikungunya virus (CHIKV). ELISA optical density was converted in Panbio units. Asterisks reflect the level of significance between groups after paired T -test was performed: ** p

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Zika virus displacement by a chikungunya outbreak in Recife, Brazil

    doi: 10.1371/journal.pntd.0006055

    Figure Lengend Snippet: Dengue virus (DENV) serological assays. (A) Amount of IgM reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (B) Amount of IgM reactive to DENV in the acute and convalescent samples of participants infected with chikungunya virus (CHIKV). (C) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with Zika virus (ZIKV). (D) Amount of IgG reactive to DENV in the acute and convalescent samples of participants infected with chikungunya virus (CHIKV). ELISA optical density was converted in Panbio units. Asterisks reflect the level of significance between groups after paired T -test was performed: ** p

    Article Snippet: CHIKV-specific IgM was assayed in the convalescent samples using an anti-chikungunya virus ELISA IgM kit (Euroimmun, Lubeck, Germany), following the manufacturer’s protocol.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay

    Numbers of CHIKV cases, anti-CHIKV IgM status and monthly precipitation in the Chapada district between October 2014 and November 2015.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Seroprevalence of Chikungunya Virus in a Rural Community in Brazil

    doi: 10.1371/journal.pntd.0005319

    Figure Lengend Snippet: Numbers of CHIKV cases, anti-CHIKV IgM status and monthly precipitation in the Chapada district between October 2014 and November 2015.

    Article Snippet: The search for anti-CHIKV IgM- and IgG-specific antibodies was performed using a commercial ELISA (enzyme-linked immunosorbent assay) according to the manufacturer's instructions (Euroimmun, Lübeck, Schleswig-Holstein, Germany).

    Techniques:

    Flow chart of the study enrollment and CHIKV IgM and IgG antibody positivity rates.

    Journal: PLoS Neglected Tropical Diseases

    Article Title: Seroprevalence of Chikungunya Virus in a Rural Community in Brazil

    doi: 10.1371/journal.pntd.0005319

    Figure Lengend Snippet: Flow chart of the study enrollment and CHIKV IgM and IgG antibody positivity rates.

    Article Snippet: The search for anti-CHIKV IgM- and IgG-specific antibodies was performed using a commercial ELISA (enzyme-linked immunosorbent assay) according to the manufacturer's instructions (Euroimmun, Lübeck, Schleswig-Holstein, Germany).

    Techniques: Flow Cytometry

    Analysis of the sensitivity and specificity of the BOB NS1 ELISA. ( A ) The values of BOB inhibition in samples from subjects diagnosed with ZIKV or DENV infection by RT-PCR are shown. The DENV group consists of samples from primary and secondary DENV cases from Nicaragua and Italy. The control group identified as “all” comprises DENV samples as well as samples from patients infected (or vaccinated) with other flaviviruses (i.e., WNV and YFV) or with CHIKV, samples from patients with other illnesses and samples from healthy blood donors. ( B ) Sensitivity calculated on the basis of different time-point cutoff values, and specificity calculated using different control groups. The flaviviruses group comprises samples from DENV and WNV cases and YFV vaccinees. The control group identified as “+ other illnesses” comprises the samples from the flavivirus group, as well as samples with CHIKV and other illnesses (Brazilian samples collected before 2014).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Antibody-based assay discriminates Zika virus infection from other flaviviruses

    doi: 10.1073/pnas.1704984114

    Figure Lengend Snippet: Analysis of the sensitivity and specificity of the BOB NS1 ELISA. ( A ) The values of BOB inhibition in samples from subjects diagnosed with ZIKV or DENV infection by RT-PCR are shown. The DENV group consists of samples from primary and secondary DENV cases from Nicaragua and Italy. The control group identified as “all” comprises DENV samples as well as samples from patients infected (or vaccinated) with other flaviviruses (i.e., WNV and YFV) or with CHIKV, samples from patients with other illnesses and samples from healthy blood donors. ( B ) Sensitivity calculated on the basis of different time-point cutoff values, and specificity calculated using different control groups. The flaviviruses group comprises samples from DENV and WNV cases and YFV vaccinees. The control group identified as “+ other illnesses” comprises the samples from the flavivirus group, as well as samples with CHIKV and other illnesses (Brazilian samples collected before 2014).

    Article Snippet: The diagnostic assessment for DENV, ZIKV, CHIKV, WNV, and YFV included the following: detection of anti-DENV IgM and IgG antibodies in serum samples (using dengue virus IgM Capture DxSelect and dengue virus IgG DxSelect, Focus Diagnostics, United States), detection of ZIKV IgM and IgG antibodies [anti-Zika virus ELISA (IgM) and anti-Zika virus ELISA (IgG); Euroimmun], identification of CHIKV IgM and IgG using specific immunofluorescence tests [anti-chikungunya virus IFA (IgG), anti-chikungunya virus IFA (IgM); Euroimmun], detection of WNV IgM and IgG using specific tests [NovaLisaTM Dengue IgM and IgG ELISAs (NovaTec Immunodiagnostic GmbH); and WNV IgM Capture DxSelect and WNV IgG DxSelect (Focus Diagnostics)], and detection of YFV IgM and IgG with anti-yellow fever virus IFA (IgG) and anti-yellow fever virus IFA (IgM; Euroimmun).

    Techniques: Enzyme-linked Immunosorbent Assay, Inhibition, Infection, Reverse Transcription Polymerase Chain Reaction

    NS1 blockade-of-binding analysis of European residents and returned travelers. ( A and B ) BOB values for samples collected in Italy, Switzerland, and the United Kingdom in ( A ) RT-PCR-confirmed ZIKV infections plotted over time. ( B ) Plotted are the BOB values in samples from ZIKV, primary and secondary DENV-, WNV-, and CHIKV-infected individuals and samples from United Kingdom-resident pregnant women from 2010 and 2015, a panel from UK individuals with symptomatic systemic illness and fever who were tested for leptospirosis (many of whom were returning travelers), and a panel of samples from healthy blood donors from Switzerland.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Antibody-based assay discriminates Zika virus infection from other flaviviruses

    doi: 10.1073/pnas.1704984114

    Figure Lengend Snippet: NS1 blockade-of-binding analysis of European residents and returned travelers. ( A and B ) BOB values for samples collected in Italy, Switzerland, and the United Kingdom in ( A ) RT-PCR-confirmed ZIKV infections plotted over time. ( B ) Plotted are the BOB values in samples from ZIKV, primary and secondary DENV-, WNV-, and CHIKV-infected individuals and samples from United Kingdom-resident pregnant women from 2010 and 2015, a panel from UK individuals with symptomatic systemic illness and fever who were tested for leptospirosis (many of whom were returning travelers), and a panel of samples from healthy blood donors from Switzerland.

    Article Snippet: The diagnostic assessment for DENV, ZIKV, CHIKV, WNV, and YFV included the following: detection of anti-DENV IgM and IgG antibodies in serum samples (using dengue virus IgM Capture DxSelect and dengue virus IgG DxSelect, Focus Diagnostics, United States), detection of ZIKV IgM and IgG antibodies [anti-Zika virus ELISA (IgM) and anti-Zika virus ELISA (IgG); Euroimmun], identification of CHIKV IgM and IgG using specific immunofluorescence tests [anti-chikungunya virus IFA (IgG), anti-chikungunya virus IFA (IgM); Euroimmun], detection of WNV IgM and IgG using specific tests [NovaLisaTM Dengue IgM and IgG ELISAs (NovaTec Immunodiagnostic GmbH); and WNV IgM Capture DxSelect and WNV IgG DxSelect (Focus Diagnostics)], and detection of YFV IgM and IgG with anti-yellow fever virus IFA (IgG) and anti-yellow fever virus IFA (IgM; Euroimmun).

    Techniques: Binding Assay, Reverse Transcription Polymerase Chain Reaction, Infection

    Diversity of the antibody response patterns in chikungunya acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and IgG (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).

    Journal: JCI Insight

    Article Title: Antibody response patterns in chikungunya febrile phase predict protection versus progression to chronic arthritis

    doi: 10.1172/jci.insight.130509

    Figure Lengend Snippet: Diversity of the antibody response patterns in chikungunya acute febrile versus chronic patients. ( A ) Paired analysis of CHIKV-specific plasma IgM (red circles) and IgG (blue squares) values in individual CHIKV-confirmed patients within the no antibodies, IgM-alone, or IgM and IgG antibody response patterns in the acute febrile (left, n = 133), early chronic (middle, n = 21), and late chronic (right, n = 30) phases. Within each antibody response pattern group, the patients are stratified based on increasing IgM values on the x axis. Horizontal dotted line indicates assay cutoff for IgM and IgG. The samples that were also positive for CHIKV PCR are indicated by green-filled symbols. ( B ) Evaluation of plasma CHIKV NT antibody activity in each of the patient groups that are described in A . Dotted gates were placed to further subgroup the patients based on a combination of IgM, IgG, and NT activity. NT assay limit of detection is indicated by the horizontal dotted line. The NT antibody titers were significantly different between the IgM-alone group and IgM and IgG group in the acute febrile patients. Statistical significance was calculated by unpaired Mann-Whitney U test. ( C ) Relative proportion of the patients with each of the indicated antibody response patterns shown in B among the CHIKV-confirmed patients in febrile phase (left), early chronic phase (middle), and late chronic phase (right).

    Article Snippet: Serology tests.CHIKV-specific IgM and IgG were detected using chikungunya IgM capture ELISA kits (Abcam ab177848) and chikungunya IgG capture ELISA kit (Abcam ab177835) following the manufacturer’s recommendations.

    Techniques: Polymerase Chain Reaction, Activity Assay, MANN-WHITNEY