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, Jose D. Debes, Khuanchai Supparatpinyo

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

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: The RT-LAMP assay detected 97.5% of HCV-RNA genotype 1, 91.1% of genotype 3, and 100% of genotype 6. The color change was evidenced with the naked eye. The assay demonstrated a clinical sensitivity of 95.5% and specificity of 100%, as well as no cross-reactivity with other viruses (i.e., hepatitis B virus, HIV). The limit of detection was as low as 10 ng per reaction for HCV genotypes 1a and 6, while it was 100 ng for genotype 3a. The assay showed a 100% detection threshold at a viral load of 5.00 log10 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.

Original languageEnglish (US)
Pages (from-to)440-445
Number of pages6
JournalInternational Journal of Infectious Diseases
Volume102
DOIs
StatePublished - Jan 2021

Bibliographical note

Publisher Copyright:
© 2020 The Author(s)

Keywords

  • Hepatitis C virus
  • Hydroxynaphthol blue
  • Loop-mediated isothermal amplification
  • Reverse transcription

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