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: International Journal of Molecular Sciences

Article Title: Evaluation of Various Alternative Economical and High Throughput SARS-CoV-2 Testing Methods within Resource-Limited Settings

doi: 10.3390/ijms232214350

Figure Lengend Snippet: Comparison of the sensitivity and specificity of the commercial SARS-CoV-2 RT-PCR assay kits.

Article Snippet: Five commercially available RT-PCR SARS-CoV-2 assay kits from different manufacturers were selected in this study for the method comparison including the Thermo Fisher TaqPathTM COVID-19 Assay Kit (Thermo Fischer Scientific, Pleasanton, CA, USA), an internationally approved kit, and the four alternative RT-PCR SARS-CoV-2 assay kits: Nucleic Acid COVID-19 Test Kit (SARS-CoV-2) (Wuhan Easy-diagnosis Biomedicine, Wuhan, China), abTES TM COVID-19 qPCR I Kit (Anatech Instrument (PTY) LTD, Meadowbrook, Business Estate, Sloane Park, Gauteng, South Africa), PCL COVID19 Speedy RT-PCR Kit (PCL Inc. Multiplex In Vitro Diagnostic Global Leader, Seoul, South Korea), and the PCLMD nCoV One-Step RT-PCR Kit (PCL Inc. Multiplex In Vitro Diagnostic Global Leader, Seoul, South Korea).

Techniques: Comparison, One Step RT-PCR