Triplex PCR Amplification of Different Melting Profile Amplicons for Simultaneous Detection of HCV, HBV, and HIV-1 Genomes in a Single Tube

Abstract

Abstract Background Hepatitis C Virus (HCV), Hepatitis B Virus (HBV), and Human Immunodeficiency Virus 1 (HIV-1) are epidemic blood-borne viruses, pose threats to human health and cause economic losses to nations for combating their infection transmission. However, the diagnostic methodologies which depend on the detection of viral nucleic acid are much expensive but more accurate than immunological testing. Methods This study approaches a lower cost triplex PCR method for simultaneous detection of the nucleic acids (genomes) of the three mentioned viruses. This PCR targets amplification of a short conserved region, having distinguishable melting profile and electrophoretic molecular weight, inside each viral genome. Therefore, this diagnostic method will be applicable with both conventional PCR (combined with electrophoresis) and real time PCR facilities. Many in silico investigations have been conducted to prove the capability of the approached PCR to detect variants of each virus. Then, Egyptian isolates of each virus have been subjected for the laboratory application of the given protocol. Results The results revealed good sensitivity for detecting and discriminating low copies of each type of viral particles. Conclusions The given protocol presents a sensitive PCR diagnostic test, with less time and cost those are ordinary needed for detecting three types of viruses. This will be a good substitutive system for the mistakable immunological-based tests.