Journal: The Lancet. Microbe

Article Title: Interference between rhinovirus and influenza A virus: a clinical data analysis and experimental infection study

doi: 10.1016/s2666-5247(20)30114-2

Figure Lengend Snippet: Data are mean (SD) of four replicates per condition. (A) Timing of sequential infections. (B) ISG mRNA expression on day 4, with or without previous rhinovirus infection. ISG expression amounts are graphed relative to the housekeeping gene HPRT . (C) Amount of IAV H1N1pdm09 viral RNA measured on days 4 and 6 with or without previous rhinovirus infection. The amount of viral RNA is expressed as fold change from the limit of detection. Significance of differences between mock pre-treated and rhinovirus pre-treated conditions were assessed by t -test (B) or two-way ANOVA (C). IAV=influenza A virus. ISG=interferon-stimulated gene. RT-qPCR=reverse-transcription quantitative PCR.

Article Snippet: To evaluate the effect of rhinovirus on subsequent infection with IAV H1N1pdm09 (strain A/California/07/2009; ATCC VR-1894), we infected differentiated airway epithelial cells with each virus individually and sequentially, then examined the time course of viral amplification and interferon-stimulated gene (ISG) induction (by RT-qPCR), using the same infection procedures as for IAV PR8-GFP.

Techniques: Expressing, Infection, Virus, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: The primers employed to detect the viral RNA in RNase digestion assay.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: The chemical structure of montelukast and its inhibitory activity against Zika virus (ZIKV) strains from Asian and African lineages in two host cells. (A) Chemical structure of montelukast. Dose-dependent inhibition of ZIKV strain SZ01 (B) , MR766 (C) , and FLR (D) infection by montelukast in BHK-21 cells and inhibition of ZIKV strain SZ01 (E) , MR766 (F) , and FLR (G) infection in Vero E6 cells. Curcumin, an anti-ZIKV drug, was included as positive control. All experiments were carried out in triplicate, and the error bars stand for standard deviation (SD). The 50% inhibitory concentration (IC50) is presented as means ± SD and summarized in .

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Activity Assay, Virus, Inhibition, Infection, Positive Control, Standard Deviation, Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: Antiviral activity of montelukast against ZIKV, DENV, and YFV in different cell lines.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Activity Assay

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: Montelukast inhibited flavivirus infection at the early stage of virus life cycle. Time of addition experiment of montelukast against Zika virus (ZIKV) SZ01 (A) , dengue virus (DENV)-2 (B) , and yellow fever virus (YFV) 17D (C) . Montelukast hardly inhibited ZIKV SZ01 (D) , DENV-2 (E) , and YFV 17D (F) infection at post-entry stage. NITD008, an adenosine analog, inhibiting flaviviruses RNA replication by terminating viral RNA synthesis was included as control. All experiments were carried out in triplicate, and the error bars stand for standard deviation (SD). The data are presented as means ± SD. NS, not significant. Student’s two-tailed t -test.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Infection, Virus, Control, Standard Deviation, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: Montelukast blocked flaviviruses adsorption, not internalization. Virus adsorption assay of montelukast against Zika virus (ZIKV) SZ01 (A) , dengue virus (DENV)-2 (B) , and yellow fever virus (YFV) 17D (C) ; and curcumin, an inhibitor known to block virus adsorption, was included as control. Virus internalization assay of montelukast against ZIKV SZ01 (D) , DENV-2 (E) , and YFV 17D (F) , and chloroquine, an inhibitor known to block virus internalization, was included as control. All experiments were carried out in triplicate, and the error bars stand for standard deviation (SD). The data are presented as means ± SD. NS, not significant; *** P < 0.001; **** P < 0.0001, Student’s two-tailed t -test.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Adsorption, Virus, Blocking Assay, Control, Standard Deviation, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: Montelukast irreversibly disrupted viral infectivity. Infectivity inhibition reversibility assay of montelukast against Zika virus (ZIKV) SZ01 (A) , dengue virus (DENV)-2 (B) , and yellow fever virus (YFV) 17D (C) . All experiments were carried out in triplicate, and the error bars stand for standard deviation (SD). The data are presented as means ± SD. NS, not significant; **** P < 0.0001, Student’s two-tailed t -test.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Infection, Inhibition, Virus, Standard Deviation, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: Degradation of released genomic RNA of flaviviruses mediated by montelukast treatment in an RNase digestion assay. The release and degradation of genomic RNA of Zika virus (ZIKV) SZ01 (A) , dengue virus (DENV)-2 (B) , yellow fever virus (YFV) 17D (C) , and purified virions of ZIKV SZ01 (D) , DENV-2 (E) , and YFV 17D (F) were detected by using their respective primers targeting different regions in the viral genome. All experiments were carried out in triplicate, and the error bars stand for standard deviation (SD). The data are presented as means ± SD.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Virus, Purification, Standard Deviation

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: Protection against vertical transmission of Zika virus (ZIKV) in montelukast-treated pregnant C57BL/6 mice. (A) Viremia of pregnant C57BL/6 mice ( n = 12 in each group). (B) Viral RNA loads in the placentas ( n = 24 in each group). Two embryos of each pregnant mouse were randomly collected, and the viral RNA load in each placenta was determined. (C) Viral RNA loads in the fetal heads ( n = 24 in each group). The viral RNA load in the fetal head of each collected embryo was determined. The bars reflect median values. The horizontal dotted lines represent limits of detection. ** P < 0.01; *** P < 0.001, Mann–Whitney test.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Transmission Assay, Virus, MANN-WHITNEY

Journal: Frontiers in Microbiology

Article Title: Montelukast, an Anti-asthmatic Drug, Inhibits Zika Virus Infection by Disrupting Viral Integrity

doi: 10.3389/fmicb.2019.03079

Figure Lengend Snippet: Protective activity of montelukast against lethal Zika virus (ZIKV) infection in type I interferon receptor-deficient A129 mice. (A) Survival of ZIKV-infected A129 mice. *** P < 0.001, log-rank (Mantel–Cox) test. (B) Viral RNA loads in sera of ZIKV-infected A129 mice. Whiskers: 5th–95th percentile. *** P < 0.001, Mann–Whitney test.

Article Snippet: ZIKV strain SZ01/2016 (GenBank number: KU866423), which was isolated from a patient who returned from Samoa and was kindly provided by Dr. Cheng-Feng Qin ( ); ZIKV strains FLR [#VR1844, American Type Culture Collection (ATCC)] and MR766 (#VR1838, ATCC) obtained from ATCC; DENV-2 kindly provided by Drs. Yunwen Hu and Zhigang Song at the Shanghai Public Health Clinical Center; and YFV strain 17D obtained from Beijing Tiantan Biological Products, Ltd., were all propagated in C6/36 cells as described previously ( ).

Techniques: Activity Assay, Virus, Infection, MANN-WHITNEY

Journal: bioRxiv

Article Title: Alignment-Free Guided Design of a Pan-Orthoflavivirus RT-qPCR Assay

doi: 10.64898/2026.03.17.712358

Figure Lengend Snippet: (A) Faceted bar charts show Ct values measured across input copy numbers (copies per µL) for each virus—dengue virus (DENV1–4), Japanese encephalitis virus (JEV), West Nile virus (WNV), yellow fever virus (YFV), and Zika virus (ZIKV). Bars summarize Ct at each concentration, with points indicating individual reactions (technical replicates); red diamonds denote the mean (error bars, where shown, indicate variability across replicates). Across DENV1–4, ZIKV, and JEV (triplicates), Ct values generally decrease as input concentration increases. ZIKV shows greater replicate-to-replicate variation at some concentrations, reflected by wider error bars compared to other targets. WNV and YFV were detected at 1,000 copies per µL; only a single concentration was tested for these viruses due to limited sample material. Together, these results indicate broad target coverage and reliable amplification across a wide input range for the main panel members. (B) Limit of detection (LOD) assessment of the designed primer-probe set. Prior to testing clinical specimens, LOD was evaluated using nucleic acids from 6 arboviruses (DENV1–4, ZIKV, and JEV) at 1, 10, 1000 copies per µL, with 12 technical replicates per dilution. The assay detected DENV3, DENV4, and JEV down to 1 copy per µL, whereas DENV1, DENV2, and ZIKV were consistently detected down to 10 copies per µL. Points represent individual reaction; a diamond indicates the mean Ct, and error bars indicate ±SD. ND indicates not detected (did not meet the ≥95% positivity criterion and therefore did not qualify as the LOD). (C) Paired comparison of Ct values between assays in clinical samples. Ct values obtained with the commercial kit (Com; circles) and the designed primer–probe set (Des; triangles) are shown for each target, with paired measurements from the same sample connected by dashed lines, for 100 positive samples and 50 positive samples (20 positive with Chikungunya virus [CHIKV] and 30 others). Box plots summarize the distributions (center line, median; box, interquartile range; whiskers, range). Relative to the commercial kit, the designed primer–probe set detected DENV1–4 significantly earlier (lower Ct; p = 0.0004 [n = 25], 0.0013 [n=25], <0.0001 [n=11] and <0.0001 [n=25] for DENV1–4, respectively) but detected ZIKV later (higher Ct; p < 0.0001 [n =14]). P values were calculated using a two-sided paired t-test on matched Com–Des Ct values for each target. Overall diagnostic performance is reported in Tables 1,2 .

Article Snippet: Reference viruses were obtained from the American Type Culture Collection (ATCC), including DENV1 strain Hawaii (ATCC VR-1856), DENV2 strain TH-36 (ATCC VR-1810), DENV3 strain H87 (ATCC VR-3380), DENV4 H241 (ATCC VR-1490), and ZIKV strain PRVABC59 (ATCC VR-1843).

Techniques: Virus, Concentration Assay, Amplification, Comparison, Diagnostic Assay