3种检测乙型肝炎病毒YMDD变异方法的比较

Comparison on detection of YMDD mutation in hepatitis B virus by three methods

目的通过多种方法比较分析找到一种高灵敏的检测乙型肝炎病毒(HBV)YMDD变异的方法。方法收集抗病毒治疗1年的慢性乙肝患者样本166例(116例采用拉米夫定治疗、50例采用替比夫定治疗),比较高温连接酶检测反应(LDR)、实时荧光聚合酶链反应(Real-time PCR)和测序3种方法检测HBV YMDD变异灵敏性。结果 LDR、Real-time PCR、测序的阳性率分别为23.49%、21.68%、19.27%。3种方法的特异性均为100%。采用拉米夫定治疗的患者中有38例变异,其中YIDD12例、YVDD26例;替比夫定只有1例YIDD变异。结论 LDR方法检测YMDD变异是非常适合临床应用的方案。YVDD变异较YIDD变异更为普遍。

Objective To analyze comparatively and find a sensitive detection method for hepatitis B virus （HBV） YMDD mutation. Methods A total of 166 samples from patients with HBV infection were collected, including 116 cases treated by lamivudine for 1 year and 50 cases treated by telbivudine for 1 year. The comparison of 3 HBV YMDD mutation detection methods real-time polymerase chain reaction （PCR）, high temperature ligase detection reaction （LDR） and sequencing was analyzed. In addition, the sensitivities of the 3 methods were detected. Results The positive rates of LDR, real-time PCR and sequencing were 23.49% , 21.68% and 19.27%. The speeificities of the 3 methods were 100%. In the cases treated by lamivudine, there were 38 cases with mutation （ 12 YIDD and 26 YVDD）. There was only 1 case with YIDD mutation in the cases treated by telbivudine. Conclusions The LDR for detecting YMDD mutation is suitable in clinical application, and YVDD mutation is more common than YIDD mutation.