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A multi‐template multiplex PCR assay for hepatitis B virus and human β‐globin
Author(s) -
Adeyemi Oluwapelumi O.,
Herod Morgan R.,
Oladiji Femi,
Fakunle Yisa M.,
Babatunde Abiola S.,
Agbede Olajide O.
Publication year - 2017
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.24877
Subject(s) - hbsag , multiplex , virology , hepatitis b virus , multiplex polymerase chain reaction , plasmid , biology , serology , microbiology and biotechnology , polymerase chain reaction , gene , antibody , virus , immunology , genetics
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 HBsAg DNA had 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.