Rapid visual detection of hepatitis C virus using a reverse transcription loop-mediated isothermal amplification assay
Sayamon Hongjaisee, Natteewan Doungjinda, Woottichai Khamduang, Tanawan Samleerat Carraway, Jiraprapa Wipasa, José D. Debes, Khuanchai Supparatpinyo
Abstract
OBJECTIVES: The aim was to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of hepatitis C virus (HCV) in a single closed tube. METHODS: Plasma samples were collected from 200 HCV-infected patients. HCV-RNA was detected by one-step RT-LAMP processed at 65 °C for 60 min. The amplified products were detected by hydroxynaphthol blue (HNB)-dependent visual method and gel electrophoresis. Specificity was tested against other viruses. Sensitivity was determined using serial dilutions of extracted RNA. RESULTS: IU/mL in the clinical samples tested. CONCLUSIONS: This study demonstrated the use of an RT-LAMP assay for the detection of HCV in a simple, rapid, and cost-effective manner, which will be useful in resource-limited settings to allow the identification of individuals in need of HCV treatment.