Published online May 28, 2016. doi: 10.3748/wjg.v22.i20.4824
Peer-review started: February 9, 2016
First decision: March 7, 2016
Revised: March 9, 2016
Accepted: March 30, 2016
Article in press: March 30, 2016
Published online: May 28, 2016
The present review describes the current status of multiplex quantitative real time polymerase chain reaction (qPCR) assays developed and used globally for detection and subtyping of hepatitis viruses in body fluids. Several studies have reported the use of multiplex qPCR for the detection of hepatitis viruses, including hepatitis A virus (HAV), hepatitis B virus (HBV), hepatitis C virus (HCV), hepatitis D virus (HDV), and hepatitis E virus (HEV). In addition, multiplex qPCR has also been developed for genotyping HBV, HCV, and HEV subtypes. Although a single step multiplex qPCR assay for all six hepatitis viruses, i.e., A to G viruses, is not yet reported, it may be available in the near future as the technologies continue to advance. All studies use a conserved region of the viral genome as the basis of amplification and hydrolysis probes as the preferred chemistries for improved detection. Based on a standard plot prepared using varying concentrations of template and the observed threshold cycle value, it is possible to determine the linear dynamic range and to calculate an exact copy number of virus in the specimen. Advantages of multiplex qPCR assay over singleplex or other molecular techniques in samples from patients with co-infection include fast results, low cost, and a single step investigation process.
Core tip: The present review describes the worldwide application and the significance of multiplex quantitative real time polymerase chain reaction (qPCR) for simultaneous detection of hepatitis viruses and their subtypes in serum. The published literature has demonstrated that the multiplex qPCR assay is a fast, easy, cost-effective, and sensitive technique for the early diagnosis of hepatitis co-infections. Use of this technique, in comparison to other diagnostic procedures, is increasing in diagnostic laboratories.