Nanoparticle-Based Lateral Flow Biosensor Integrated With Loop-Mediated Isothermal Amplification for Rapid and Visual Identification of Chlamydia trachomatis for Point-of-Care Use

• Published in: Frontiers in microbiology Vol. 13; p. 914620
• Main Authors: Chen, Xu, Zhou, Qingxue, Tan, Yan, Wang, Ronghua, Wu, Xueli, Liu, Jiangli, Liu, Rui, Wang, Shuoshi, Dong, Shilei
• Format: Journal Article
• Language: English
• Published: Frontiers Media S.A 12-07-2022
• Subjects: Chlamydia trachomatis; gold nanoparticle-based lateral flow biosensor; limit of detection; loop-mediated isothermal amplification; Microbiology; point-of-care testing
• ISSN: 1664-302X; 1664-302X
• DOI: 10.3389/fmicb.2022.914620

Chlamydial infection, caused by
Chlamydia trachomatis
, is the most common bacterial sexually transmitted infection and remains a major public health problem worldwide, particularly in underdeveloped regions. Developing a rapid and sensitive point-of-care (POC) testing for accurate screening of
C. trachomatis
infection is critical for earlier treatment to prevent transmission. In this study, a novel diagnostic assay, loop-mediated isothermal amplification integrated with gold nanoparticle-based lateral flow biosensor (LAMP-LFB), was devised and applied for diagnosis of
C. trachomatis
in clinical samples. A set of LAMP primers based on the
ompA
gene from 14
C. trachomatis
serological variants (serovar A-K, L1, L2, L3) was successfully designed and used for the development of
C. trachomatis
-LAMP-LFB assay. The optimal reaction system can be performed at a constant temperature of 67°C for 35 min. The total assay process, including genomic DNA extraction (~15 min), LAMP reaction (35 min), and LFB readout (~2 min), could be finished within 60 min. The
C. trachomatis
-LAMP-LFB could detect down to 50 copies/ml, and the specificity was 100%, no cross-reactions with other pathogens were observed. Hence, our
C. trachomatis
-LAMP-LFB was a rapid, reliable, sensitive, cost-effective, and easy-to-operate assay, which could offer an attractive POC testing tool for chlamydial infection screening, especially in resource starvation settings.