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: 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: Compared with the qPCR technology ( C. trachomatis real-time TaqMan PCR method, DaAn Gene Co., Ltd. China), the MCDA-AuNPs-LFB sensitivity, specificity, positive predictive value, and negative predictive value were 100%, 96.20%, 94.92%, and 100%, respectively ( ).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

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: Our assay was robustly assessed using suspected C. trachomatis -infection genital secretion samples and compared with a commercial C. trachomatis real-time TaqMan PCR kit (DaAn Gene Co., Ltd. China).

Techniques:

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Visual and rapid identification of Chlamydia trachomatis and Neisseria gonorrhoeae using multiplex loop-mediated isothermal amplification and a gold nanoparticle-based lateral flow biosensor

doi: 10.3389/fcimb.2023.1067554

Figure Lengend Snippet: Schematic diagram showing AuNPs-LFB principles for visual mLAMP amplification product interpretation (A) mLAMP amplification products (0.5 μL) and running buffer (100 μL) were simultaneously added to the sample pad. (B) The running buffer containing the mLAMP products moves forward onto the conjugate pad and nitrocellulose membrane by capillary action. SA-AuNPss are hydrated, rapidly released, and combined with C . trachomatis or N. gonorrhoeae LAMP products at the conjugate pad. (C) FAM/biotin-labeled C . trachomatis LAMP products are captured by anti-FAM at TL1, Dig/biotin-labeled N. gonorrhoeae LAMP products are captured by anti-Dig at TL2, and SA-AuNPs are captured by biotin-BSA at the CL. (D) Interpretation of the mLAMP-AuNPs-LFB assay. C . trachomatis positive results are indicated by CL and TL1 bands on the AuNPs-LFB, N. gonorrhoeae positive results are indicated by CL and TL2 bands on the AuNPs-LFB, and C . trachomatis and N. gonorrhoeae positive results are indicated by TL1, TL2, and CL bands on the AuNPs-LFB. Negative results are indicated when only the CL band appears on the AuNPs-LFB. Key: CL, control line; TL1, test line one; TL2, test line two; CT, C . trachomatis ; NG, N. gonorrhoeae ; NC, nitrocellulose membrane.

Article Snippet: We compared the mLAMP-AuNPs-LFB assay to commercially available real-time TaqMan PCR Kits for C. trachomatis and N. gonorrhoeae (DaAn Gene Co., Ltd.; Guangzhou, China) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies; Singapore).

Techniques: Amplification, Membrane, Labeling, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Visual and rapid identification of Chlamydia trachomatis and Neisseria gonorrhoeae using multiplex loop-mediated isothermal amplification and a gold nanoparticle-based lateral flow biosensor

doi: 10.3389/fcimb.2023.1067554

Figure Lengend Snippet: mLAMP product confirmation and verification The mLAMP products were simultaneously identified using (A) malachite green and (B) the AuNPs-LFB. Tube/AuNPs-LFB 1 shows a positive C . trachomatis result. Tube/AuNPs-LFB 2 shows a positive N. gonorrhoeae result. Tube/AuNPs-LFB 3 shows a positive C . trachomatis and N. gonorrhoeae result. Tube/AuNPs-LFB 4 shows negative control (DW). Key: CL, control line; TL1, test line one; TL2, test line two; CT, C . trachomatis ; NG, N. gonorrhoeae .

Article Snippet: We compared the mLAMP-AuNPs-LFB assay to commercially available real-time TaqMan PCR Kits for C. trachomatis and N. gonorrhoeae (DaAn Gene Co., Ltd.; Guangzhou, China) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies; Singapore).

Techniques: Negative Control, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Visual and rapid identification of Chlamydia trachomatis and Neisseria gonorrhoeae using multiplex loop-mediated isothermal amplification and a gold nanoparticle-based lateral flow biosensor

doi: 10.3389/fcimb.2023.1067554

Figure Lengend Snippet: Temperature optimization for the ( C ) trachomatis and N. gonorrhoeae LAMP amplification LAMP amplifications for (A) C . trachomatis and (B) N. gonorrhoeae were monitored using real-time turbidity, and their corresponding amplicon curves are shown as graphs. A turbidity >0.1 indicated a positive result. Eight kinetic graphs were obtained at different temperatures (62°C–69°C in 1°C increments) with 1×10 4 target gene copies. Graphs e (66°C) to h (69°C) in (A) showed robust amplification. Graphs from f (67°C) to h (69°C) in (B) showed robust amplification.

Article Snippet: We compared the mLAMP-AuNPs-LFB assay to commercially available real-time TaqMan PCR Kits for C. trachomatis and N. gonorrhoeae (DaAn Gene Co., Ltd.; Guangzhou, China) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies; Singapore).

Techniques: Amplification

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Visual and rapid identification of Chlamydia trachomatis and Neisseria gonorrhoeae using multiplex loop-mediated isothermal amplification and a gold nanoparticle-based lateral flow biosensor

doi: 10.3389/fcimb.2023.1067554

Figure Lengend Snippet: Sensitivity analysis of the mLAMP-AuNPs-LFB assay with serial nucleic acid template dilutions Serial dilutions (5.0×10 4 , 5.0×10 3 , 5.0×10 2 , 5.0×10 1 , 5.0×10 0 , and 5.0×10 -1 copies) of C. trachomatis ompA and N. gonorrhoeae orf1 plasmids were used as templates, and DW was used as the negative control. Results were simultaneously analyzed by visual reagent malachite green and the AuNPs-LFB. (A, B) : A sensitivity analysis of the C. trachomatis LAMP assay indicated its LoD was 50 copies per reaction. (C, D) : A sensitivity analysis of the N. gonorrhoeae LAMP assay indicated its LoD was 50 copies per reaction. (E, F) : A sensitivity analysis of the mLAMP assay for ompA and orf1 indicated its LoD was 50 copies of the nucleic acid template per reaction. Key: CL, control line; TL1, test line one; TL2, test line two; CT, C. trachomatis ; NG, N. gonorrhoeae ; MG, malachite green.

Article Snippet: We compared the mLAMP-AuNPs-LFB assay to commercially available real-time TaqMan PCR Kits for C. trachomatis and N. gonorrhoeae (DaAn Gene Co., Ltd.; Guangzhou, China) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies; Singapore).

Techniques: Negative Control, Lamp Assay, Mlamp Assay, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Visual and rapid identification of Chlamydia trachomatis and Neisseria gonorrhoeae using multiplex loop-mediated isothermal amplification and a gold nanoparticle-based lateral flow biosensor

doi: 10.3389/fcimb.2023.1067554

Figure Lengend Snippet: mLAMP-AuNPs-LFB assay specificity with different strains Assay specificity was evaluated using different nucleic acid templates. Amplification products were tested using AuNPs-LFBs: 1–14, C. trachomatis serovars A, B, C, D, E, F, G, H, I, J, K, L1, L2, and L3 ompA plasmids; 15–20, C. trachomatis clinical samples; 21, N. gonorrhoeae orf1 plasmids; 22, N. gonorrhoeae reference strain ATCC 49926; 23–28, N. gonorrhoeae clinical samples; 29–34, C. trachomatis and N. gonorrhoeae clinical samples; 35, Neisseria meningitides ; 36, Ureaplasma urealyticum ; 37, Escherichia coli ; 38, Staphylococcus aureus ; 39, human papillomavirus; 40, Mycoplasma pneumonia ; 41, Haemophilus influenza ; 42, Streptococcus pyogenes ; 43, human enterovirus EV71; 44, Coxsackie virus CAV16; 45, Klebsiella pneumoniae ; 46, Pseudomonas aeruginosa ;47, Candida glabrata ; 48, Cryptococcus neoformans ; 49, Listeria monocytogenes ; 50, negative control (DW). Key: CL, control line; TL, test line.

Article Snippet: We compared the mLAMP-AuNPs-LFB assay to commercially available real-time TaqMan PCR Kits for C. trachomatis and N. gonorrhoeae (DaAn Gene Co., Ltd.; Guangzhou, China) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies; Singapore).

Techniques: Amplification, Virus, Negative Control, Control

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Visual and rapid identification of Chlamydia trachomatis and Neisseria gonorrhoeae using multiplex loop-mediated isothermal amplification and a gold nanoparticle-based lateral flow biosensor

doi: 10.3389/fcimb.2023.1067554

Figure Lengend Snippet: Comparing C. trachomatis and N. gonorrhoeae levels in clinical samples using our mLAMP-AuNPs-LFB assay and qPCR method.

Article Snippet: We compared the mLAMP-AuNPs-LFB assay to commercially available real-time TaqMan PCR Kits for C. trachomatis and N. gonorrhoeae (DaAn Gene Co., Ltd.; Guangzhou, China) on an Applied BiosystemsTM 7500 Real-Time PCR System (Life Technologies; Singapore).

Techniques: