Affiliation:
1. The Second Clinical Medical College Guizhou University of Traditional Chinese Medicine Guiyang Guizhou People's Republic of China
2. Department of Scientific Research, The Second Affiliated Hospital Guizhou University of Traditional Chinese Medicine Guiyang Guizhou People's Republic of China
3. Central Laboratory of the Second Affiliated Hospital Guizhou University of Traditional Chinese Medicine Guiyang Guizhou People's Republic of China
4. Department of Clinical Laboratory Zhejiang Hospital Hangzhou Zhejiang People's Republic of China
5. Experimental Center Guizhou Provincial Center for Disease Control and Prevention Guiyang Guizhou People's Republic of China
6. Department of Gastroenterology, The Second Affiliated Hospital Guizhou University of Traditional Chinese Medicine Guiyang Guizhou People's Republic of China
7. Clinical Laboratory Hangzhou Women′s Hospital Hangzhou Zhejiang People's Republic of China
Abstract
AbstractHepatitis C remains a global health problem, especially in poverty‐stricken areas. A rapid and sensitive point‐of‐care (POC) diagnostic tool is critical for the early detection and timely treatment of hepatitis C virus (HCV) infection. Here, for the first time, we reported a novel molecular diagnostic assay, termed reverse transcription multiple cross displacement amplification integrated with a gold‐nanoparticle‐based lateral flow biosensor (RT‐MCDA‐AuNPs‐LFB), which was developed for rapid, sensitive, specific, and visual identification of HCV. HCV‐RT‐MCDA induced rapid isothermal amplification through a specific primer set targeting the 5′untranslated region gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a that are prevalent in China. The optimal reaction temperature and time for RT‐MCDA‐AuNPs‐LFB were 68°C and 25 min, respectively. The limit of detection of the assay was 10 copies per test, and the specificity was 100% for the experimental strains. The whole detection procedure, including crude nucleic acid isolation (~5 min), RT‐MCDA (68°C, 25 min), and visual AuNPs‐LFB result confirmation (less than 2 min), was performed within 35 min. The preliminary results indicated that the HCV‐RT‐MCDA‐AuNPs‐LFB assay could be a valuable tool for sensitive, specific, visual, cost‐saving, and rapid detection of HCV and has potential as a POC diagnostic platform for field screening and early clinical detection of HCV infection.
Funder
Medical Scientific Research Foundation of Zhejiang Province, China