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[Quantitative analysis of HBV DNA amplified products with microtiter hybridization]
Quan Zhang1, Jing-juan Ding, Xiao-hui Miao
1Department of Infection Disease, Guiyang Medical College, Guizhou 550004, China.
Background:
To establish a new assay for detecting the quantity of HBV DNA with PCR and enzyme-linked immunosorbent assay(ELISA).
Methods:
The 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.
Results:
Totally 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.
Conclusions:
The PCR ELISA assay is simple and suitable for clinical laboratory in quantitative determination of HBV DNA.