|
Objective To detect hepatitis B virus (HBV) YMDD mutation of three kinds of methods for comparing the sensitivity and specificity, combined with patients before treatment HBV DNA level, ALT levels and immune markers and YMDD mutation HBVDNA load and other clinical information analysis of results. Methods to fluorescence quantitative PCR (FQ-PCR), the generic template signal amplification (UT-PCR) and polymerase chain reaction microplate hybridization - enzyme-linked immunosorbent assay (PCRmnh-ELISA), lamivudine treatment serum HBV DNA by the overcast sun appeared for 52 patients with chronic hepatitis B patients for HBV YMDD mutation detection, comparing the sensitivity of the three methods, and the results of the three methods detect inconsistent samples were sequenced, comparison of their specificity . While the pre-treatment HBV DNA level, ALT levels, immune markers and HBV DNA level mutation YMDD mutation results combined with a comprehensive analysis. Results FQ-PCR, UT-PCR and PCRmnh-ELISA for the detection of YMDD mutation rates were: 53.85%, 48.08%, 73.07%, the pairwise comparison, FQ-PCR and UT-PCR were associated with significantly PCRmnh-ELISA difference (P <0.05), while the FQ-PCR and UT-PCR was no significant difference (P> 0.1). The test results are inconsistent 17 specimens were sequenced, FQ-PCR, UT-PCR, PCRmnh-ELISA and sequencing results of the compliance rate was 94.1%, 70.6%, 29.4%, there was significant difference between the three groups (P < 0.005). 52 cases of patients, 28 cases occurred in YMDD mutations, which YIDD 17 例, YVDD 7 例, YIDD and YVDD hybrid mutation in 4 cases. YIDD mutation occurred before treatment in patients with HBV DNA levels were significantly lower (P <0.05), occurred YVDD variation than before treatment in patients with HBV DNA levels had no significant difference (P> 0.05). YVDD mutated HBVDNA levels and HBV DNA levels YIDD mutation was no significant difference (P> 0.05). YMDD mutation before treated with baseline ALT levels above the level of YMDD wild group before treatment (P <0.01), pre-treatment HBV DNA levels and HBeAg positive rate of mutation and wild group undifferentiated group (P> 0.05). Conclusion ① 3 Ways comparison, FQ-PCR detection of HBV YMDD mutation has a high sensitivity and specificity, and can simultaneously detect the wild-type strain and mutants in mixed infections, and sequencing results consistent with the diagnosis of HBV YMDD mutation better Methods. UT-PCR compared with PCRmnh-ELISA, high specificity, and can specifically detect YVDD, YIDD two kinds of variation, and the sequencing results meet higher. These results suggest that, FQ-PCR and UT-PCR method in the detection of HBV YMDD mutation superior PCRmnh-ELISA method, worthy of clinical application. ② PCRmnh-ELISA as cumbersome operation and high false positive rate, it is proposed methodology is improved before considering clinical applications. ③ lamivudine therapy based on high levels of ALT may be a YMDD mutation prone factors. Closely monitored in patients with chronic hepatitis HBV DNA and ALT levels, whether it is conducive to timely detection of YMDD mutation. ④ dynamic monitoring of HBV YMDD variation, during lamivudine therapy helps to detect YMDD mutation, timely adjustment of treatment.
|