Rapid Visual Detection of Hepatitis C Virus Using Reverse Transcription Recombinase-Aided Amplification-Lateral Flow Dipstick.

Haili Wang,Xifang Zhu,Peiyang Ding,Yuhang Zhang,Yumei Chen,Chao Liang,Aiping Wang,Ying Zhang,Ming Li,Hongliang Liu,Yankai Liu,Jingming Zhou,Cheng Xin,Gaiping Zhang

doi:10.3389/fcimb.2022.816238

Abstract

Hepatitis C virus (HCV) infection is a global public health threat. Reaching the World Health Organization’s objective for eliminating viral hepatitis by 2030 will require a precise disease diagnosis. While immunoassays and qPCR play a significant role in detecting HCV, rapid and accurate point-of-care testing is important for pathogen identification. This study establishes a reverse transcription recombinase-aided amplification–lateral flow dipstick (RT-RAA-LFD) assay to detect HCV. The intact workflow was completed within 30 min, and the detection limit for synthesized C/E1 plasmid gene-containing plasmid was 10 copies/μl. In addition, the test showed good specificity, with no cross-reactivity observed for hepatitis A virus, hepatitis B virus, HIV, syphilis, and human papillomavirus virus. Using extracted RNAs from 46 anti-HCV antibody-positive samples, RT-RAA-LFD showed 100% positive and negative concordance rates with qPCR. In summary, the RT-RAA-LFD assay established in this study is suitable for the rapid clinical detection of HCV at the community level and in remote areas.

Highlights

Infectious diseases pose a critical global threat that has caused significant morbidity and mortality as well as great economic losses (Houghton, 2009; Massimo and Vincenzo, 2018)
Hepatitis C virus (HCV) infection is a major cause of severe liver diseases, including chronic hepatitis, cirrhosis, and hepatocellular carcinoma
According to the World Health Organization (WHO, 2017), approximately 71 million people worldwide are infected with HCV, and at least 400,000 people die annually from HCV-related liver disease (WHO, 2017; Spearman et al, 2019; Roger et al, 2021)

Summary

Introduction

Infectious diseases pose a critical global threat that has caused significant morbidity and mortality as well as great economic losses (Houghton, 2009; Massimo and Vincenzo, 2018). Hepatitis C virus (HCV) infection is a major cause of severe liver diseases, including chronic hepatitis, cirrhosis, and hepatocellular carcinoma. According to the World Health Organization (WHO, 2017), approximately 71 million people worldwide are infected with HCV, and at least 400,000 people die annually from HCV-related liver disease (WHO, 2017; Spearman et al, 2019; Roger et al, 2021). Acute HCV cases account for about 15–20% of total cases, and post-acute HCV-infected patients have a 50–80% chance of. Rapid Visual Detection of HCV developing a chronic infection (Narayanamurthy et al, 2021). Patients have a 20% risk of developing cirrhosis, of whom a small percentage may develop hepatocellular carcinoma (Roger et al, 2021)

Methods

Results

Conclusion