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

Frontiers in Cellular and Infection Microbiology

Cellular and Infection Microbiology

Qingxue Zhou¹  Shilei Dong²  Xinhua Luo³  Wei Yuan⁴  Yuanfang Shi⁵  Xu Chen⁶ 

[1]Clinical Laboratory, Hangzhou Women’s Hospital, Hangzhou, Zhejiang, China

[2]Department of Clinical Laboratory, Zhejiang Hospital, Hangzhou, Zhejiang, China

[3]Department of Infectious Disease, Guizhou Provincial People’s Hospital, Guiyang, Guizhou, China

[4]Department of Quality Control, Guizhou Provincial Center for Clinical Laboratory, Guiyang, Guizhou, China

[5]The Second Clinical College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, China

[6]The Second Clinical College, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, China

[7]Clinical Medical Laboratory of the Second Affiliated Hospital, Guizhou University of Traditional Chinese Medicine, Guiyang, Guizhou, China

关键词: Chlamydia trachomatis;    Neisseria gonorrhoeae;    loop-mediated isothermal amplification;    gold nanoparticle-based lateral flow biosensor;    point-of-care testing;   

DOI  :  10.3389/fcimb.2023.1067554

 received in 2022-10-12, accepted in 2023-02-15,  发布年份 2023

来源: Frontiers

【 摘 要 】

Sexually transmitted chlamydia and gonorrhea infections caused by the bacteria Chlamydia trachomatis and Neisseria gonorrhoeae remain a major public health concern worldwide, particularly in less developed nations. It is crucial to use a point of care (POC) diagnostic method that is quick, specific, sensitive, and user-friendly to treat and control these infections effectively. Here, a novel molecular diagnostic assay, combining multiplex loop-mediated isothermal amplification (mLAMP) with a visual gold nanoparticles-based lateral flow biosensor (AuNPs-LFB) was devised and used for highly specific, sensitive, rapid, visual, and easy identification of C. trachomatis and N. gonorrhoeae. Two unique independent primer pairs were successful designed against the ompA and orf1 genes of C. trachomatis and N. gonorrhoeae, respectively. The optimal mLAMP-AuNPs-LFB reaction conditions were determined to be 67°C for 35 min. The detection procedure, involving crude genomic DNA extraction (~5 min), LAMP amplification (35 min), and visual results interpretation (<2 min), can be completed within 45 min. Our assay has a detection limit of 50 copies per test, and we did not observe any cross-reactivity with any other bacteria in our testing. Hence, our mLAMP-AuNPs-LFB assay can potentially be used for POC testing to detect C. trachomatis and N. gonorrhoeae in clinical settings, particularly in underdeveloped regions.

【 授权许可】

Unknown   
Copyright © 2023 Chen, Zhou, Yuan, Shi, Dong and Luo