Application of a NASBA Real-time assay using molecular beacon for detection of HCV virus

Background and ObjectivesHepatitis C is the most common viral disease among intravenous drug users transmitted mainly through blood transfusion. In this study, we used an isothermal nucleic acid sequence-based amplification (NASBA) in combination with a molecular beacon probe-based real-time assay for detection of HCV. Materials and MethodsIn this experimental study, the conserved 5’NCR region with the length of 241 bp was used for probe and primers design. In comparison with the standard, RNA virus detection sensitivity of the method was 100 percent for up to 500 copies/ ml. ResultsNo serological positive sample was detected with this assay; the clinical sensitivity of reaction was 96.6%. NCBI Nucleotide Blast showed 100% analytical specificity and no cross was observed with viruses or human genome. Investigation of 10 serological negative samples showed the clinical specificity to be 100%. ConclusionsIn summary, due to its isothermal nature, its speed, and its use for RNA specific amplification, NASBA real-time assay will have broad applications for the rapid detection of HCV in plasma samples.