应用实时荧光PCR方法检测乙型肝炎病毒基因YMDD变异

Applicate method of real time PCR to detect YMDD variation in HBV

[目的]建立一种检测乙型肝炎患者YMDD变异株的方法,并且研究应用拉米夫定治疗及未用药治疗的乙型肝炎初发患者中YMDD变异株发生的情况。[方法]应用实时荧光聚合酶链反应(PCR)方法对30例未经过任何药物治疗的乙肝初发患者,50例使用拉米夫定治疗1年患者,50例使用拉米夫定治疗两年患者和20例使用拉米夫定与干扰素联合治疗两年的患者同时进行YMDD变异株的检测。[结果]通过对熔解曲线的分析,在未经任何治疗的乙肝初发患者中检出YMDD变异株2例,占6.67%;拉米夫定治疗1年的患者中检出YMDD变异株13例,占26.00%;拉米夫定治疗两年患者中检出YMDD变异株25例,占50.00%;拉米夫定和干扰素联合治疗的患者中检出YMDD变异株9例,占45.00%。[结论]在未经任何药物治疗的乙肝患者中即存在YMDD变异株,并且随着使用拉米夫定时间的延长在不断增加,且联合用药并不能减少变异株的发生,因此对乙肝患者在用药前,及用药过程中进行YMDD变异株的检测是非常必要的。而实时荧光PCR方法具有简便、快速、灵敏、经济的特点,特别适用于临床对YMDD变异株的检测,从而及时的指导临床用药。

[ Objective] Setting up a method to detect YMDD mutation during lamivudine therapy and study the application pull the rice man settles the treatment and did not use the initial stages in hepatitis in type in B of the medicine treatment the sufferer the circumstance of the inside YMDD variation stub occurrence, [ Methods] The application comes together with the real time PCR method to the early hair in liver in B for having not yet any medicine treatment sufferer,50 usages pulls the rice man settles the sufferer who cure a year,50 usages pulls the sufferer that rice man settle two years in treatment to pull with 20 usages the rice man settle with the intederon sufferer of uniting two years in treatment at the same time the examination that proceed the YMDD variation stub. [Results ] Pass to solve the curvilinear analysis to the liquate linear , examine the 2 examples ：YMDD variation stub in earlier in liver in B that had not yet any treatment sufferer, occupy 6,67% ;Pulling the rice man settle the sufferer who cure to examine the 13 examples ：YMDD variation stub in a year, occupying 26, 00% ; Pulling the rice man settles two years in treatment to examine the 25 examples ： YMDD variation stub in the sufferer, occupying 50, 00%, pulling the rice man settle to examine the 9 examples ： YMDD variation stub with interferon in the sufferer of uniting the treatment, occupying 45, 00%. [ Conclusion] Exsits namely in had not yet any medicine treatment of B liver sufferer YMDD variation stub, and pull along with the usage the rice man settles the horary extension at increase continuously, and in association with use the medicine and cant reduce the occurrence of the variation stub, therefore to B liver the sufferer proceeds in use medicine front, and use medicine process the YMDD variation stub examination is very necessary, The method described here provides a convenient,rapid ,accurate and inexpensive way to monitor YMDD mutation during lamivudine therapy.