Biotechnology and Applied Biochemistry · 2020 · 24 citations · 10 references
MedicineInfectious DiseasesMolecular BiologyMolecular DiagnosticsMolecular VirologyVirologyMolecular ScreeningNucleic Acid Amplification TestingPathogen DetectionPolymerase Chain ReactionViral DiagnosticsViral HepatitisAmplificationDiagnostic Assay DevelopmentHepatitis Virus DiagnosisHepatitis Virus TestingPolymerase Chain Reaction (Pcr) AmplificationPolymerase Chain Reaction-based DiagnosticsRecombinase Polymerase AmplificationSerum Sampling
Hepatitis B virus (HBV) is a worldwide epidemic pathogen that causes hepatitis B. On-site screening the HBV infection is of critical importance for preventing and diagnosing HBV infection. In this paper, a simple, visual, and rapid method for on-site detection of HBV-DNA has been developed. This method is based on betaine-assisted recombinase polymerase assay and followed with naked-eye detection via lateral flow assay (BRPA-LF). Result show that nonspecific amplification is prone to occur in recombinase polymerase amplification (RPA) if the assay was performed with serum sample without purification. This problem has been addressed by adding 0.8 M of betaine to the RPA reactions. It was demonstrated that BRPA-LF can detect 1,000 copies of HBV-DNA in 50 μL mixture, and achieved 90% sensitivity and 100% specificity for serum sample detection. These results demonstrated that BRPA-LF can resist serum interference and has great potential for on-site screening of HBV infection.
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