title: Multiplex qPCR for serodetection and serotyping of hepatitis viruses: A brief review.
abstract: 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.
authors.affiliation: Mohammad Irshad, Priyanka Gupta, Dhananjay Singh Mankotia, Mohammad Ahmad Ansari, Clinical Biochemistry Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi 110029, India.
authors.forename: Mohammad
authors.initials: M
authors.lastname: Irshad
authors.affiliation: Mohammad Irshad, Priyanka Gupta, Dhananjay Singh Mankotia, Mohammad Ahmad Ansari, Clinical Biochemistry Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi 110029, India.
authors.forename: Priyanka
authors.initials: P
authors.lastname: Gupta
authors.affiliation: Mohammad Irshad, Priyanka Gupta, Dhananjay Singh Mankotia, Mohammad Ahmad Ansari, Clinical Biochemistry Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi 110029, India.
authors.forename: Dhananjay Singh
authors.initials: DS
authors.lastname: Mankotia
authors.affiliation: Mohammad Irshad, Priyanka Gupta, Dhananjay Singh Mankotia, Mohammad Ahmad Ansari, Clinical Biochemistry Division, Department of Laboratory Medicine, All India Institute of Medical Sciences, New Delhi 110029, India.
authors.forename: Mohammad Ahmad
authors.initials: MA
authors.lastname: Ansari
mesh_terms: D004279:DNA, Viral / Q000097:blood / Q000235:genetics*; D006518:Hepatitis Viruses / Q000145:classification / Q000235:genetics* / Q000276:immunology; D006525:Hepatitis, Viral, Human / Q000097:blood / Q000175:diagnosis* / Q000235:genetics / Q000276:immunology; D006801:Humans; D060885:Multiplex Polymerase Chain Reaction; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D065288:Serogroup; D012698:Serologic Tests / Q000379:methods*; D012703:Serotyping
chemical_list: D004279:DNA, Viral
keywords: Co-infection; Genotyping techniques; Hepatitis viruses; Quantitative real-time polymerase chain reaction; Serotyping; Viral genome