Application Of Real-Time Fluorescence Quantitative PCR In The Laboratory Diagnosis For Chronic Hepatitis B Cured With IFN

Objective To investigate the value of real-time fluorescence quantitative PCR (FQ-PCR) used in the predication and evaluation of curative effect of interferon (IFN) for chronic hepatitis B(CHB).Methods 45 cases of CHB patients were divided into two groups, therapy and control, randomly. The cases of therapeutic group were cured with IFN and common support therapy while the control group was treated only with the common support means. The period of treatment was 6 months, and after that they had been traced for an additional 12 months. The blood samples were taken at 9 time points, i. e, pretreatment, 1,2,3,4,5,6 months after the therapy was done, and 6,12 months after the IFN treatment finished. Hepatitis B virus DNA (HBV-DNA) was quantified with the FQ-PCR; HBeAg was quantitatively detected by the sandwich assay of radial immunity. The other serum markers of HBVXHBsAg, anti-HBs, anti-HBc, anti-HBe) were also qualified with the same method as used in HBeAg.Results (1) HBV-DNA in the therapeutic group decreased gradually in the first 5 months after IFN was used. (P<0. 01),and it had diminished from 107.57 to 104.97 in that period, but it didn't fall down after that. The HBV-DNA remained at this low level for the 12 months' following up . (2) There were good correlations between HBV-DNA and HBeAg, HBV-DNA and ALT in the therapeutic group before treatment. (r = 0. 719 and 0. 492) . However, negative correlations were found between HBV-DNA and ALT at every time points of treatment and tracing, bad correlations were found between HBV-DNA and HBeAg in the same period(r<0. 2) . (3)HBV-DNA in the therapeutic group was thd same as that in the control group before treatment. Although the HBV-DNA in the two groups had fallen down month by month, no significant difference was found in the first 5 month period. (P>0. 05) between the two groups, while the HBV-DNA was lower in the therapeutic group(104.90 copies/ml) than that of control groupdO5.52 copies/ml) at the time point of the sixth month after curing (P<0. 05). Seven cases in the therapeutic group developed a complete response after treatment(33. 3%) , and HBsAg in one case among them Rversed had been found negative reversion negative. reverse (5%), and two cases ex-pressed part-response(9%) . The total rate of effect was 42%. While a complete response was found only in 3 cases in the control group (12. 5%), and they all turned positive at the time point of 12 months' following-up. HBV-DNA in the patients who expressed a complete response wasl06.51 copies, which is lower than that of those who had were no response (107.72 copies/ml) in the therapeutic group. (P<0.01).Conclusion The cases had to be selected carefully depending on the copies of HBV-DNA and other related factors before IFN was used in curing CHB in order to get better result and avoid the side effect and financial waste caused by IFN misused. Up until now, the FQ-PCR was the first choice that was the most sensitive, convenient, accurate. and timesaving way in HBV-DNA quantity. When the IFN curing effect for CHB was evaluated, FQ-PCR was more sensitive and accurate than the factors of HBeAg and ALT, furthermore, it could find out the cases whose HBVMwere negative while the HBV-DNA was positive. The more reliable reference will be obtained when the 3 factors are used comprehensively.