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: 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:

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effects of 405 nm irradiance on chlamydial growth in HeLa cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5. ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (24 h post). Treatments are grouped based on post-hoc comparisons for convenience. The effect of 405 nm on chlamydial growth was assessed during active and persistent stages induced with penicillin ( B and C ). Growth was determined using quantitative real-time PCR to determine the ratio of chlamydial and eukaryotic housekeeping genes (16S: GAPDH respectively) 48 h post-infection on cDNA reverse transcribed from RNA. Mean ± standard deviation are plotted for the two replicated experiments. Statistical significance was determined post-hoc using a Bonferonni adjustment comparing all groups against C. trachomatis -infected HeLa cells alone (CTE); * P < 0.05, ** P < 0.005.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Real-time Polymerase Chain Reaction, Reverse Transcription, Standard Deviation

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Anti-chlamydial properties of 405 nm irradiance. ( A - C ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 without exposure to photodiodes. ( D - F ) Infected cells were exposed to 405 nm LEDs at 20 J/cm 2 promptly after infection to evaluate anti-chlamydial effects during an acute chlamydial infection. Cells were fixed and stained with dapi (blue) ( B and E ) and anti-chlamydial (green) ( C and F ) antibody 48 hours post-infection. Bar = 10μm.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Staining

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effect of 405 nm on IL-6 production in C. trachomatis -infected epithelial cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 (CTE5). ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (post-24 h). The effect of 405 nm on IL-6 production was assessed during active ( A and B ) and penicillin-induced persistent stages ( C ). Supernatants were collected and measured for IL-6 production using an ELISA. Treatments are grouped based on post-hoc comparisons for convenience. Mean ± SEM are plotted for the two replicated experiments. Statistical differences were determined post-hoc using a Bonferonni adjustment comparing all groups to C. trachomatis infected cells (CTE); *, P < 0.05; ** P < 0.005.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal: BMC Microbiology

Article Title: Inhibitory effects of 405 nm irradiation on Chlamydia trachomatis growth and characterization of the ensuing inflammatory response in HeLa cells

doi: 10.1186/1471-2180-12-176

Figure Lengend Snippet: Effect of 405 nm on CCL2 production in C. trachomatis -infected epithelial cells. ( A ) HeLa cells were infected with C. trachomatis serovar E at a MOI of 5 (CTE5). ( B ) Infected cells were then exposed to varying doses of 405 nm at a range of energy densities (5-20 J/cm 2 ) either promptly after infection or 24 h post-infection (post-24 h). The effect of 405 nm on CCL2 was assessed during active ( B ) and persistent stages induced with penicillin ( C ). Supernatants were collected and measured for CCL2 production using an ELISA. Treatments are grouped based on post-hoc comparisons for convenience. Mean ± SEM are plotted for the two replicated experiments. Statistical differences were determined post-hoc using a Bonferonni adjustment comparing all groups to C. trachomatis infected cells (CTE); *, P < 0.001.

Article Snippet: C. trachomatis -infected HeLa cells with or without 405 nm were fixed with ice-cold methanol for 10 min. After aspiration, culture wells were washed with PBS and then stained with rabbit anti- C. trachomatis EBs (Virostat, Portland, ME) for 1 h. Wells were washed five times with PBS and counterstained with 4’, 6-diamidino-2’-phenylindole, dihydrochloride (Dapi; Thermo Scientific, Rockford, IL) for 10 min.

Techniques: Infection, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Measurement of IgG antibodies to Chlamydia trachomatis by commercial enzyme immunoassays and immunofluorescence in sera from pregnant women and patients with infertility, pelvic inflammatory disease, ectopic pregnancy, and laboratory diagnosed Chlamydia psittaci / Chlamydia pneumoniae infection

doi:

Figure Lengend Snippet: Specificity of the commercial enzyme immunoassays (EIAs) for detecting Chlamydia trachomatis antibody when tested against a panel of Chlamydia psittaci/Chlamydia pneumoniae antibody positive sera as diagnosed by whole cell immunofluorescence assay

Article Snippet: Sera for testing were diluted 1/64 in DELFIA assay buffer (Wallac Oy, Turku, Finland) and 100 μl was loaded on to Genzyme Virotech C trachomatis coated microtitre plates (Genzyme Virotech).

Techniques: Enzyme Immunoassay, Immunofluorescence

Journal: BMC Infectious Diseases

Article Title: Do host genetic traits in the bacterial sensing system play a role in the development of Chlamydia trachomatis -associated tubal pathology in subfertile women?

doi: 10.1186/1471-2334-6-122

Figure Lengend Snippet: The pattern recognition receptors (PRRs), which recognize C. trachomatis pathogen-associated molecular patterns (PAMPs), and the single nucleotide polymorphisms (SNPs) studied

Article Snippet: In our hands, the C. trachomatis IgG titre obtained by the C. pneumoniae MIF (AniLabsystems) had the best predictive value for tubal factor subfertility [ ].

Techniques:

Journal: BMC Infectious Diseases

Article Title: Do host genetic traits in the bacterial sensing system play a role in the development of Chlamydia trachomatis -associated tubal pathology in subfertile women?

doi: 10.1186/1471-2334-6-122

Figure Lengend Snippet: The risk of tubal pathology (TP) in C. trachomatis IgG-positive subfertile women in relation to the genotype of the single pattern recognition receptor genes. a Adapted from Morré et al ., 2003 [1]. b Adapted from Ouburg et al ., 2005 [2].

Article Snippet: In our hands, the C. trachomatis IgG titre obtained by the C. pneumoniae MIF (AniLabsystems) had the best predictive value for tubal factor subfertility [ ].

Techniques:

Journal: BMC Infectious Diseases

Article Title: Do host genetic traits in the bacterial sensing system play a role in the development of Chlamydia trachomatis -associated tubal pathology in subfertile women?

doi: 10.1186/1471-2334-6-122

Figure Lengend Snippet: The risk of tubal pathology (TP) in C. trachomatis IgG-positive (CT+) and IgG-negative (CT-) subfertile women in relation to carrying five single nucleotide polymorphisms (SNPs) in four pattern recognition receptor genes.

Article Snippet: In our hands, the C. trachomatis IgG titre obtained by the C. pneumoniae MIF (AniLabsystems) had the best predictive value for tubal factor subfertility [ ].

Techniques: