A multi-template multiplex PCR assay for hepatitis B virus and human β-globin
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Date
2017
Authors
Adeyemi, Oluwapelumi
Herod, Morgan
Oladiji, Femi
Fakunle, Yisa
Babatunde, Abiola
Agbede, Olajide
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Publisher
Wiley-Blackwell, USA
Abstract
The Hepatitis B surface antigen (HBsAg) is the hallmark of HBV infection. Detection of antibodies to HBs and the core (ie, 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 β-globin is a well characterized housekeeping gene (HKG) that is often applied as internal controls (IC) in molecular diagnosis. In this study, individual plasmid clones of the human β-globin and HBs genes were constructed. These plasmid constructs have been applied to characterize a multiplex PCR assays for HBs and β-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 β-globin and HBsAgDNAhad a resultant effect on assay sensitivity. This study further highlights the importance of molecular diagnosis in HBV infectious
individuals. If fully optimized, this assay could provide a possible diagnostic complement to serological detection in developing countries.
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Citation
Adeyemi O.O., Herod M.R., Oladiji F., Fakunle Y.M., Babatunde A.S., and 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