Rapid detection of hepatitis C virus using recombinase polymerase amplification

Area of Science:

• Virology
• Molecular Diagnostics
• Public Health

Background:

• Hepatitis C virus (HCV) infects over 71 million globally, causing significant mortality from untreated chronic infections.
• Direct-acting antivirals (DAAs) offer high cure rates but require accurate HCV RNA diagnostics for patient identification.
• Current PCR-based tests are centralized, limiting access to rapid diagnosis in diverse clinical settings.

Purpose of the Study:

• To develop and evaluate a rapid, minimally instrumented assay for Hepatitis C virus RNA detection.
• To assess the assay's performance across all HCV genotypes and its clinical accuracy.
• To explore the potential of this assay for point-of-care implementation to improve global access to HCV treatment.

Main Methods:

• Development of a reverse transcription recombinase polymerase amplification (RT-RPA) assay for HCV RNA detection.
• Evaluation of the assay's limit of detection for genotype 1 HCV RNA.
• Clinical validation using 78 diverse serum specimens to determine sensitivity and specificity.

Main Results:

• The RT-RPA assay detected all HCV genotypes with a limit of detection of 25 copies/reaction for genotype 1.
• Achieved 100% clinical sensitivity and 100% specificity in evaluating 78 clinical serum specimens.
• Demonstrated rapid performance with low heating requirements, suitable for decentralized testing.

Conclusions:

• The RT-RPA assay provides a rapid, accurate, and accessible method for Hepatitis C virus RNA detection.
• Its minimal instrumentation and high performance support potential implementation as a point-of-care test.
• This advancement can significantly expand global access to timely chronic HCV diagnosis and DAA treatment.