Quantitative analysis of HBV DNA amplified products with microtiter hybridization.
- Author:
Quan ZHANG
1
;
Jing-juan DING
;
Xiao-hui MIAO
Author Information
- Publication Type:Journal Article
- MeSH: DNA Probes; DNA, Viral; analysis; biosynthesis; genetics; Enzyme-Linked Immunosorbent Assay; Hepatitis B; virology; Hepatitis B Antibodies; blood; Hepatitis B Surface Antigens; blood; Hepatitis B e Antigens; blood; Hepatitis B virus; genetics; isolation & purification; Humans; Polymerase Chain Reaction; Sensitivity and Specificity
- From: Chinese Journal of Experimental and Clinical Virology 2003;17(1):39-41
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUNDTo establish a new assay for detecting the quantity of HBV DNA with PCR and enzyme-linked immunosorbent assay(ELISA).
METHODSThe products of PCR using primers pre-labeled with biotin were hybridized with the capture probes that were immobilized on the microtiter strips and then bond with Sav-Ap. The quantity of DNA was detected by measuring the yellow color at 405 nm wave length.
RESULTSTotally 125 sera from patients with hepatitis B were tested for HBV DNA by this method,the sera were also tested for HBV immunological markers by solid phase radio immuno-assay (SPRIA). The HBV positive rate with PCR-ELISA was 64.9% (24/37) in samples which were positive for HBsAg, HBeAg and HBcAb; and 34.2% (13/38) in sera which were positive for HBsAg, HBeAb and HBcAb; in sera positive for HBsAg and HBcAb or only HBcAb, the positive rate was 6.7% (1/15) and 5.9% (2/34) respectively.
CONCLUSIONSThe PCR ELISA assay is simple and suitable for clinical laboratory in quantitative determination of HBV DNA.
