Quantification of hepatitis B virus DNA using competitive PCR and a scintillation proximity assay.
A rapid assay for the quantification of hepatitis B virus DNA in human serum was developed. The principle of the method combines competitive polymerase chain reaction (cPCR) (for the controlled amplification of hepatitis B virus DNA) and scintillation proximity assay (SPA) technology (for rapid detection and quantitation of PCR products). It also incorporates a reproducible and simple method for the preparation of serum DNA suitable for PCR amplification. The assay has a better linear dynamic range than traditional methods that use 32P to detect PCR products. It was applied to a range of hepatitis B virus (HBV) surface antigen positive (HBsAg + ) sera, and shown to be more sensitive than a commercially available HBV DNA kit.
Simpson
P R
PR
Scitech Genetics Ltd., The Maxwell, Singapore, Singapore.
Yu
X H
XH
Redza
Z M
ZM
Anson
J G
JG
Chan
S H
SH
Lin
Y
Y
D004279:DNA, Viral / Q000097:blood*; D003947:Diagnostic Techniques, Radioisotope; D005784:Gene Amplification; D006509:Hepatitis B / Q000821:virology*; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification / Q000502:physiology*; D006801:Humans; D010761:Phosphorus Radioisotopes; D016133:Polymerase Chain Reaction; D011933:Reagent Kits, Diagnostic; D015203:Reproducibility of Results; D012588:Scintillation Counting; D012680:Sensitivity and Specificity; D019562:Viral Load
D004279:DNA, Viral; D010761:Phosphorus Radioisotopes; D011933:Reagent Kits, Diagnostic