A multi-Template Multiplex PCR Assay for Hepatitis B virus and Human β-globin
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Date
2017
Journal Title
Journal ISSN
Volume Title
Publisher
Wiley
Abstract
The Hepatitis B surface antigen (HBsAg) is the hallmark of HBV infection. Detection of
antibodies to HBs and the core (i.e. HBsAg and HBcAb) are primary serological algorithms in
the laboratory diagnosis of HBV. Detection of HBsAg DNA is an important supplement to
serological diagnosis especially in clinical cases. Simultaneous amplification of internal
cellular controls is a good indicator of sample quality. Human P-globin is a well characterised
housekeeping gene (HKG) that is often applied as internal controls (IC) in molecular
diagnosis. In this study, individual plasmid clones of the human P-globin and HBs genes were
constructed. These plasmid constructs have been applied to characterise a multiplex PCR
assays for HBs and P-globin genes. The findings suggest detection limits of less than 10
genome copies of either template In vitro using conventional and multiplex PCR conditions.
Under the multiplex conditions, co-amplification of P-globin and HBsAg DNA had a resultant
effect on assay sensitivity. This study further highlights the importance of molecular diagnosis
in HBV infectious individuals. If fully optimised, this assay could provide a possible
diagnostic complement to serological detection in developing countries.
Description
Keywords
Hepatitis B,, HBs gene,, Housekeeping, Multiplex Polymerase Chain Reaction,
Citation
Adeyemi, O.O., Herod, M.R., Oladiji, F., Fakunle, Y.M., Babatunde, A.S., & Agbede, O.O. (2017). A multi-Template Multiplex PCR Assay for Hepatitis B virus and Human β-globin. Journal of Medical Virology, 89(11): 1944 -1951.