Quantitative analysis of HBV DNA amplified products with microtiter hybridization / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>BACKGROUND</b>To establish a new assay for detecting the quantity of HBV DNA with PCR and enzyme-linked immunosorbent assay(ELISA).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>The PCR ELISA assay is simple and suitable for clinical laboratory in quantitative determination of HBV DNA.</p>