Journal: PLoS Pathogens

Article Title: Attachment and Entry of Chlamydia Have Distinct Requirements for Host Protein Disulfide Isomerase

doi: 10.1371/journal.ppat.1000357

Figure Lengend Snippet: CHO6 cells were transfected with vectors expressing wildtype PDI (CHO6+PDI) or an enzymatic mutant of PDI lacking active site cysteine residues (CHO6+PDI-4CS). 48 h after transfection, cells were infected with Chlamydia for subsequent attachment and entry analysis. Cells were fixed and evaluated by immunofluorescence, bacteria are green and counter staining shown with Evans blue. (A) Bacterial attachment was analyzed 1 h post-infection. Bacterial attachment was recovered in CHO6+PDI as well as in CHO6+PDI-4CS, indicating that PDI enzymatic activity is not necessary for bacterial attachment. (B) The number of bacteria attached to cells was determined by quantification, the number of bacteria associated with each cell in eight separate fields of view containing at least ten cells. Error bars indicate standard of deviation (STDEV). (C) 24 h after infection Chlamydia infectivity was evaluated. Infectivity was restored in CHO6+PDI. No productive infection was observed in CHO6 cells or in CHO6+PDI-4CS. In CHO6+PDI-4CS, the bacteria remained persistently attached to the cell, indicative of the requirement for PDI enzymatic activity for bacterial entry. (D) Infection was quantified by counting the number of inclusions per field of view in eight separate fields of view containing at least ten cells. Error bars indicate the STDEV. (E) Cells were incubated at 37°C for 2 h to allow for bacterial entry. Entry was analyzed by comparing the total number of cell-associated bacteria for staining of permeabilized cells (Total) and the number of extracellular bacteria determined by staining unpermeabilized cells (Extracellular). Bacterial entry was recovered in CHO6 cells expressing PDI (CHO6+PDI) but not in CHO6 cells expressing enzymatically nonfunctional PDI (CHO6+PDI-4CS). (F) Percent internalization represents 1 minus the number of extracellular bacteria divided by the total number of bacteria multiplied by 100. The number of extracellular and total bacteria was determined by quantifying the number of bacteria associated with each cell per field of view in eight separate fields of view containing at least ten cells. Error bars indicate the STDEV.

Article Snippet: Blocking solution was removed and coverslips were incubated 1 h with mouse anti- Chlamydia MOMP antibody for C. trachomatis or anti- Chlamydia LPS antibody (Santa Cruz Biotechnology, Santa Cruz, CA) for C. psittaci and then washed with HBSS.

Techniques: Transfection, Expressing, Mutagenesis, Infection, Immunofluorescence, Bacteria, Staining, Activity Assay, Standard Deviation, Incubation

Journal: PLoS Pathogens

Article Title: Attachment and Entry of Chlamydia Have Distinct Requirements for Host Protein Disulfide Isomerase

doi: 10.1371/journal.ppat.1000357

Figure Lengend Snippet: (A) Chlamydia attachment to CHOK1 was evaluated by immunofluorescence staining of bacteria (green) 1 h after infection and counter staining with Evans blue. Cells were infected and then maintained in media containing 0 mM or 3 mM bacitracin. Similar levels of bacterial attachment were observed with and without bacitracin. (B) Entry was examined after a 2 h incubation at 37°C. Entry was analyzed by comparing the total number of cell-associated bacteria (permeabilized) and the number of extracellular bacteria (unpermeabilized). 3 mM bacitracin inhibited bacterial entry. (C) The effect of bacitracin on Chlamydia attachment and infection was quantified and is shown as the % attachment and infection of cells not treated with bacitracin. Attachment was analyzed 1 h post-infection and infection was evaluated 24 h post-infection. (D) The effect of bacitracin on Chlamydia development was examined by evaluating cells 24 h post-infection that had not been treated with bacitracin (a′), been treated with 3 mM bacitracin for 20 min prior to infection (b′), been treated with bacitracin for the first 8 h of infection (c′), or been treated with bacitracin only for the last 16 h of the 24 h infection (d′). In all cases the development of bacteria-containing vacuoles was seen, indicative of a normal infection.

Article Snippet: Blocking solution was removed and coverslips were incubated 1 h with mouse anti- Chlamydia MOMP antibody for C. trachomatis or anti- Chlamydia LPS antibody (Santa Cruz Biotechnology, Santa Cruz, CA) for C. psittaci and then washed with HBSS.

Techniques: Immunofluorescence, Staining, Bacteria, Infection, Incubation

Journal: PLoS Pathogens

Article Title: Attachment and Entry of Chlamydia Have Distinct Requirements for Host Protein Disulfide Isomerase

doi: 10.1371/journal.ppat.1000357

Figure Lengend Snippet: (A) CHO6 cells transfected with a vector expressing PDI with a gpi-anchor (PDI-gpi) are marked with a white arrow, untransfected cells are indicated by a yellow arrow. Cells were fixed and stained for PDI using PDI-specific antibody. Expression of PDI-gpi led to a large amount of PDI localized at the cell surface. (B) Cell survival was analyzed 72 h after DT treatment. CHOK1 cells were extremely sensitive to toxin, whereas CHO6 cells were relatively resistant. Expression of PDI-gpi in CHOK1 or CHO6 cells did not alter toxin sensitivity, indicative of an inability of PDI-gpi to properly interact with other cell surface proteins. (C) Chlamydia attachment to CHOK1 and CHO6+PDI-gpi was analyzed, bacteria are shown in green, and PDI is stained red. There was no bacterial attachment to CHO6+PDI-gpi.

Article Snippet: Blocking solution was removed and coverslips were incubated 1 h with mouse anti- Chlamydia MOMP antibody for C. trachomatis or anti- Chlamydia LPS antibody (Santa Cruz Biotechnology, Santa Cruz, CA) for C. psittaci and then washed with HBSS.

Techniques: Transfection, Plasmid Preparation, Expressing, Staining, Bacteria

Journal: PLoS Pathogens

Article Title: Attachment and Entry of Chlamydia Have Distinct Requirements for Host Protein Disulfide Isomerase

doi: 10.1371/journal.ppat.1000357

Figure Lengend Snippet: (A) Immunoblot of cells transfected with PDI-targeted siRNA or non-targeting siRNA using antibody to PDI. GAPDH expression was evaluated to ensure equivalent loading. (B) Following siRNA treatment, C. psittaci and C. trachomatis attachment were examined. PDI staining is shown in red, and bacteria are stained green. Because of the significant PDI downregulation, cells in PDI siRNA panels are marked by arrows. siRNA downregulation of PDI led to a dramatic decrease in bacterial attachment, but attachment was not affected by treatment with non-targeting siRNA. (C) The number of bacteria attached to cells following treatment with non-targeting siRNA (gray) or PDI siRNA (black) was determined by quantification of the number of cell-associated bacteria by counting eight separate fields of view containing at least ten cells. Error bars indicate standard error of the mean (SEM).

Article Snippet: Blocking solution was removed and coverslips were incubated 1 h with mouse anti- Chlamydia MOMP antibody for C. trachomatis or anti- Chlamydia LPS antibody (Santa Cruz Biotechnology, Santa Cruz, CA) for C. psittaci and then washed with HBSS.

Techniques: Western Blot, Transfection, Expressing, Staining, Bacteria

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Genetic Inactivation of Chlamydia trachomatis Inclusion Membrane Protein CT228 Alters MYPT1 Recruitment, Extrusion Production, and Longevity of Infection

doi: 10.3389/fcimb.2018.00415

Figure Lengend Snippet: Histopathological assessment of reproductive tracts post-infection. Mice ( n = 5 per group) were euthanized at 23 and 64 days post-infection (dpi) and entire reproductive tracts were removed and formalin fixed for histology. Representative images of H&E stained sections of uterine tissue were captured using an Olympus DP70 for mice infected with (A) C. trachomatis L2-wild type, 23 dpi, (B) C. trachomatis L2-ΔCT228, 23 dpi, (C) C. trachomatis L2-wild type, 64 dpi, (D) C. trachomatis L2-ΔCT228, 64 dpi. See Table for pathological clinical scoring of reproductive tracts at 64 dpi.

Article Snippet: Chlamydia trachomatis serovar L2-wild type and L2-ΔCT228 (LGV 434/Bu, originating from the Hackstadt Lab at Rocky Mountain Laboratories, Hamilton, MT) were propagated in HeLa 229 cells and purified by Renografin density gradient centrifugation as previously described (Caldwell et al., ).

Techniques: Infection, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Genetic Inactivation of Chlamydia trachomatis Inclusion Membrane Protein CT228 Alters MYPT1 Recruitment, Extrusion Production, and Longevity of Infection

doi: 10.3389/fcimb.2018.00415

Figure Lengend Snippet: TargeTron inactivation of CT228 . (A) TargeTron insertion site in CT228 . (B) Schematic of CT228 containing GII( aadA ). (C) PCR verification of TargeTron insertion in L2-ΔCT228. PCRs were used to amplify incA, CT228 and the Intron in the L2-wild type, L2-ΔCT228 (Mut), and TargeTron vector (pDFTT295). (D) Immunofluorescence images of L2-wild type and L2-ΔCT228 at 18 h post-infection stained with anti-CT228 and anti- Chlamydia LPS followed by fluorescent secondary antibodies. Scale bar, 10 μm.

Article Snippet: Chlamydia trachomatis serovar L2-wild type and L2-ΔCT228 (LGV 434/Bu, originating from the Hackstadt Lab at Rocky Mountain Laboratories, Hamilton, MT) were propagated in HeLa 229 cells and purified by Renografin density gradient centrifugation as previously described (Caldwell et al., ).

Techniques: Plasmid Preparation, Immunofluorescence, Infection, Staining

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Genetic Inactivation of Chlamydia trachomatis Inclusion Membrane Protein CT228 Alters MYPT1 Recruitment, Extrusion Production, and Longevity of Infection

doi: 10.3389/fcimb.2018.00415

Figure Lengend Snippet: TargetTron inactivation of Chlamydia trachomatis CT228 and subsequent CT229-CT224 gene linkage group transcription. (A) Physical map of C. trachomatis CT229-CT224 gene linkage group. Solid arrows represent CT229-CT224 ORFs. Gene designations are indicated above each ORF. Green and red arrows indicate forward and reverse primers, respectively, used for RT-PCR analysis. Red arrowhead indicates the point of TargeTron insertion within CT228 . (B) Agarose gel image(s) show RT-PCR products corresponding to groEL (control), and CT229-CT224 gene linkage group expression (correlating to the indicated primers above). Total RNA (L2-wild type or L2-ΔCT228) is indicated above corresponding gel lanes. + indicates RT added to reaction. – Indicates RT not added to reaction. L2-wild type and L2-ΔCT228 DNA act as PCR controls. (C) Growth curves of L2-wild type and L2-ΔCT228 were performed in triplicate, the entire experiment was repeated on three separate occasions and a representative growth curve was selected. Error bars represent standard deviation.

Article Snippet: Chlamydia trachomatis serovar L2-wild type and L2-ΔCT228 (LGV 434/Bu, originating from the Hackstadt Lab at Rocky Mountain Laboratories, Hamilton, MT) were propagated in HeLa 229 cells and purified by Renografin density gradient centrifugation as previously described (Caldwell et al., ).

Techniques: Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Expressing, Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Genetic Inactivation of Chlamydia trachomatis Inclusion Membrane Protein CT228 Alters MYPT1 Recruitment, Extrusion Production, and Longevity of Infection

doi: 10.3389/fcimb.2018.00415

Figure Lengend Snippet: Recruitment of MYPT1 and Myosin phosphatase pathway components and extrusion production by C. trachomatis L2-wild type and L2-ΔCT228. HeLa cell monolayers were infected at a MOI of ~0.5 with L2-wild type and L2-ΔCT228 for 18 h (in technical triplicate). Cells were fixed and stained with primary antibodies to MYPT1, Chlamydia LPS, MLC2 (pS19), Src Y474, MLCK (pY471), non-muscle Myosin IIa and IIb followed by fluorescent secondary antibodies. Experiments were repeated on three separate occasions and representative images were selected. (A,B) Top panel shows individual and merged images of MYPT1 recruitment (green) and Chlamydia LPS staining (red) in both the L2-wild type and L2-ΔCT228. Lower panel of individual and merged images show MLC2 (pS19), MLCK (pY471), and Mysoin IIa and IIb (green) co-localizing with active Src Y474 kinase (red) in microdomains at the periphery of inclusions in both L2-wild type and L2-ΔCT228. Scale bar, 10 μm. (C) Total protein from L2-wild type and L2-ΔCT228 infected HeLa cells at 24 and 48 h post-infection were assessed for MLC2, MLC2 (pS19), HsP60, and GAPDH levels by western blot analysis. (D) HeLa cells were treated with either Scramble (Scr) or MYPT1 siRNA for 48 h prior to infection with L2-wild type and L2-ΔCT228. Protein samples were assessed for MYPT1 and GAPDH levels by western blot. (E) Extrusions collected and (F) IFUs were assessed for L2 wild-type and L2-ΔCT228 at 48 h post-infection in either Scramble (symbols and solid bars) or MYPT1 (open symbols and white bars) siRNA treated HeLa cells. * p < 0.0001.

Article Snippet: Chlamydia trachomatis serovar L2-wild type and L2-ΔCT228 (LGV 434/Bu, originating from the Hackstadt Lab at Rocky Mountain Laboratories, Hamilton, MT) were propagated in HeLa 229 cells and purified by Renografin density gradient centrifugation as previously described (Caldwell et al., ).

Techniques: Infection, Staining, Western Blot

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Genetic Inactivation of Chlamydia trachomatis Inclusion Membrane Protein CT228 Alters MYPT1 Recruitment, Extrusion Production, and Longevity of Infection

doi: 10.3389/fcimb.2018.00415

Figure Lengend Snippet: Recoverable IFUs shed by mice infected with C. trachomatis L2-wild type and L2-ΔCT228. Female C3H/HeJ mice were intravaginally infected with 1 × 10 6 EBs of either L2-wild type or L2-ΔCT228. Recoverable IFUs were obtained by swabbing vaginal tracts and enumerating on HeLa cell monolayers. Recoverable IFU data are expressed for (A) L2-wild type ( n = 8) and (B) L2-ΔCT228 ( n = 9) on a logarithmic scale from Day 7 to 42 post-infection. Effect of time ( * p = 0.006) was observed in mice infected with L2-wild type (repeated measures one-way ANOVA). For day 28, 6/8 L2-wild type infected mice were clear compared to 3/9 L2- ΔCT228. Triangles represent individual mice, bars represent mean of group for each time point.

Article Snippet: Chlamydia trachomatis serovar L2-wild type and L2-ΔCT228 (LGV 434/Bu, originating from the Hackstadt Lab at Rocky Mountain Laboratories, Hamilton, MT) were propagated in HeLa 229 cells and purified by Renografin density gradient centrifugation as previously described (Caldwell et al., ).

Techniques: Infection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Genetic Inactivation of Chlamydia trachomatis Inclusion Membrane Protein CT228 Alters MYPT1 Recruitment, Extrusion Production, and Longevity of Infection

doi: 10.3389/fcimb.2018.00415

Figure Lengend Snippet: Systemic and mucosal antibody titers following infection with C. trachomatis L2-wild type and L2-ΔCT228. (A) Sera were collected from mice 31 days post-infection with L2-wild type ( n = 8) or L2-ΔCT228 ( n = 9) and assayed for the presence of anti-chlamydial IgG2a. (B,C) Vaginal lavages were collected 31 days post-infection and assayed for the presence of anti-chlamydial IgA and IgG. Antibody titers are expressed for individual mice as the highest dilution tested that produced >3-fold the absorbance as control/uninfected mice. Bars represent mean antibody titer per group. * p = 0.0412, unpaired two-tailed Student t -test.

Article Snippet: Chlamydia trachomatis serovar L2-wild type and L2-ΔCT228 (LGV 434/Bu, originating from the Hackstadt Lab at Rocky Mountain Laboratories, Hamilton, MT) were propagated in HeLa 229 cells and purified by Renografin density gradient centrifugation as previously described (Caldwell et al., ).

Techniques: Infection, Produced, Two Tailed Test

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Genetic Inactivation of Chlamydia trachomatis Inclusion Membrane Protein CT228 Alters MYPT1 Recruitment, Extrusion Production, and Longevity of Infection

doi: 10.3389/fcimb.2018.00415

Figure Lengend Snippet: Pathological scoring of murine reproductive tracts.

Article Snippet: Chlamydia trachomatis serovar L2-wild type and L2-ΔCT228 (LGV 434/Bu, originating from the Hackstadt Lab at Rocky Mountain Laboratories, Hamilton, MT) were propagated in HeLa 229 cells and purified by Renografin density gradient centrifugation as previously described (Caldwell et al., ).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: C . trachomatis -MCDA-AuNPs-LFB assay workflow. The workflow includes genomic DNA preparation, MCDA amplification, and AuNP-LFB visual interpretation, all completed within 40 min.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Amplification

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: Schematic diagram showing AuNPs-LFB principles for the visual identification of C trachomatis -MCDA amplification products. (A) C trachomatis -MCDA amplification products (0.5 μl) and running buffer (100 μl) were simultaneously added to the sample pad. (B) Due to capillary action, the running buffer, containing (C) trachomatis -MCDA products, moved forward onto the conjugate pad and nitrocellulose (NC) membrane. Streptavidin-AuNPs were hydrated, rapidly released, and combined with C trachomatis -MCDA products at the conjugate pad. (C) FAM/biotin-labeled C trachomatis -MCDA products were arrested by anti-FAM at the TL strip, and streptavidin-DPNs were arrested at the biotin-BSA CL strip. (D) Interpretation of the C trachomatis -AuNP-LFB assay. For a positive result, both the CL and TL appeared on the biosensor. For a negative result, only the CL was observed on the AuNP-LFB. TL: test line; CL: control line.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Amplification, Membrane, Labeling, Stripping Membranes, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: C . trachomatis -MCDA-AuNPs-LFB degenerate primers used in this study.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Sequencing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: Confirmation and verification of (C) trachomatis -MCDA products. C trachomatis -MCDA products were measured simultaneously using malachite green (MG) (A) and AuNPs-LFB (B) . Tube 1/Biosensor 1: positive result for C trachomatis ompA standard plasmids; Tube 2/Biosensor 2: negative result for Neisseria gonorrhoeae ; Tube 3/Biosensor 3: negative result for Ureaplasma urealyticum ; Tube 4/Biosensor 4: blank control (distilled water, DW). TL: test line; CL: control line.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: Optimizing the temperature for the C. trachomatis -MCDA assay. C. trachomatis -MCDA amplification of ompA was monitored using real-time turbidity. Corresponding amplicon concentration curves are marked in graphs. Turbidity > 0.1 indicated a positive value. (A–H) Eight kinetic graphs were generated at different temperatures (63°C–70°C at 1°C intervals) with C. trachomatis ompA -plasmids at 1 × 10 3 copies. Graph E (67°C) showed the fastest and most robust amplification.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Amplification, Concentration Assay, Generated

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: Sensitivity analysis of C trachomatis -MCDA-AuNPs-LFB using C trachomatis ompA -plasmid serial dilutions. Serial dilutions (1.0 × 10 4 , 1.0 × 10 3 , 1.0 × 10 2 , 1.0 × 10 1 , 1.0 × 10 0 , and 1.0 × 10 −1 copies) of C trachomatis ompA -plasmids were used as templates, and distilled water (DW) was used as the negative control. Results were simultaneously analyzed by malachite green (MG) (A) and AuNPs-LFB (B) . The limit of detection (LoD) for C trachomatis -MCDA-AuNP-LFB was 10 copies/test. CL, control line; TL, test line.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Plasmid Preparation, Negative Control, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: Optimal amplification time for the C. trachomatis -MCDA-AuNPs-LFB assay. Four reaction times ( A , 10 min; B , 20 min; C , 30 min; and D , 40 min) were evaluated at 67°C. Tubes/biosensors 1–7 represented C. trachomatis ompA template levels: 1.0 × 10 4 , 1.0 × 10 3 , 1.0 × 10 2 , 1.0 × 10 1 , 1.0 × 10 0 , 1.0 × 10 −1 copies, and negative control (distilled water, DW), respectively. Results were simultaneously analyzed using malachite green (MG) and AuNP-LFB. The optimal limit of detection (LoD) occurred when the amplification lasted for 30 min (C) . CL: control line; TL: test line.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Amplification, Negative Control, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: Analytical specificity of the C. trachomatis -MCDA-AuNPs-LFB assay using different strains. Assay specificity was evaluated using different nucleic acids as temperatures, and products were tested using AuNPs-LFB. Biosensors 1–14, C. trachomatis serovars A, B, C, D, E, F, G, H, I, J, K, L1, L2, and L3 ompA -plasmids; Biosensors 15–21, C. trachomatis (clinical samples); Biosensor 22, Ureaplasma urealyticum ; Biosensor 23, Neisseria gonorrhoeae ; Biosensor 24, Escherichia coli ; Biosensor 25, Staphylococcus aureus ; Biosensor 26, Human papilloma virus; Biosensor 27, Human rhinovirus; Biosensor 28, Coxsackie virus CAV16; Biosensor 29, Human enterovirus EV71; Biosensor 30, Mycoplasma pneumoniae ; Biosensor 31, Listeria monocytogenes ; Biosensor 32, Haemophilus influenza ; Biosensor 33, Cryptococcus neoformans ; Biosensor 34, Bordetella pertussis ; Biosensor 35, Streptococcus pyogenes ; Biosensor 36, Candida glabrata ; Biosensor 37, Pseudomonas aeruginosa ; Biosensor 38, Shigella flexneri ; Biosensor 39, Klebsiella pneumoniae ; Biosensor 40, negative control (distilled water, DW). CL: control line; TL: test line.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: Virus, Negative Control, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Sensitive and visual identification of Chlamydia trachomatis using multiple cross displacement amplification integrated with a gold nanoparticle-based lateral flow biosensor for point-of-care use

doi: 10.3389/fcimb.2022.949514

Figure Lengend Snippet: Comparing C. trachomatis levels in clinical samples using our MCDA-AuNPs-LFB assay with a qPCR method.

Article Snippet: Using 135 suspected C. trachomatis -infected genital secretion samples from Hangzhou Women’s Hospital (Hangzhou, China), we compared our assay with a commercial C. trachomatis real-time TaqMan PCR method (DaAn Gene Co., Ltd. China) (Cat. #DA0071) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies, Singapore), which was used as a reference method since it is commonly used in clinical Chinese laboratories, and its sensitivity was verified using C. trachomatis standard substance (Guangzhou BDS Biological Technology Co., Ltd.).

Techniques: