A multi-template multiplex PCR assay for hepatitis B virus and human β-globin.
Oluwapelumi Olufemi AdeyemiMorgan R HerodFemi OladijiYisa M FakunleAbiola S BabatundeOlajide O AgbedePublished in: Journal of medical virology (2017)
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.
Keyphrases
- hepatitis b virus
- real time pcr
- high throughput
- endothelial cells
- liver failure
- genome wide
- label free
- escherichia coli
- induced pluripotent stem cells
- nucleic acid
- loop mediated isothermal amplification
- pluripotent stem cells
- machine learning
- gene expression
- deep learning
- wastewater treatment
- cell free
- single cell
- quantum dots
- mass spectrometry
- copy number
- quality improvement
- simultaneous determination
- bioinformatics analysis