Concurrent quantitation of the A and D genotypes of hepatitis B virus.
Hepatitis B virus (HBV) infection is a global health problem associated with severe liver disorders. Viral load and HBV genotype affect the clinical outcome, guide antiviral therapy and provide long term prognosis for HBV infected patients. Various types of detection and quantitation assays are currently in use with a different effectiveness. The aim of this study was to develop a method that would provide simultaneous identification and quantitation of genotypes A and D in a single-tube reaction. Sera from infected patients were analyzed by a TaqMan based real time PCR. Optimized reagents were used for HBV DNA quantitation while the genotypes A and D were quantified separately by our design of the assay. Multiplex real time PCR was achieved and was specific for HBV genotypes A and D within a single-tube reaction. Simulation of mixed virus populations was identified reproducibly in vitro. Quantitation of these individual genotypes was exceptionally reliable, so much so that the sum of individual genotypes was equal to the total viral load determined in a separate reaction. Therefore, a straightforward, conceptually simple and reliable approach to issues involving HBV genotypes A and D is submitted. Identity and exact titer of these genotypes in the Caucasian population can now be determined easily.
Tanic
Nikola
N
Department of Neurobiology, Institute for Biological Research Sinisa Stanković, University of Belgrade, Belgrade, Serbia. nikolata@ibiss.bg.ac.rs
Stanojevic
Boban
B
Tanic
Nasta
N
Schaefer
Stephan
S
Niesters
Hubert G M
HG
Bozic
Milena
M
Dimitrijevic
Bogomir
B
D001483:Base Sequence; D004279:DNA, Viral / Q000032:analysis / Q000097:blood / Q000235:genetics; D014644:Genetic Variation; D006515:Hepatitis B virus / Q000235:genetics* / Q000302:isolation & purification; D019694:Hepatitis B, Chronic / Q000097:blood / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D019562:Viral Load; D044465:White People
D004279:DNA, Viral