荧光分子信标探针检测拉米夫定耐药基因的研究

Clinical and Experimental Study on Lumivudine Resistance by Molecular Beacon

建立荧光分子信标探针方法 ,探讨乙肝病毒对拉米夫定耐药的发生及病程进展。观察 77例乙肝病人口服拉米夫定治疗前及治疗后 1 2个月及 2 4个月肝功能 (ALT)及免疫学指标及检测HBVDNA含量 ,并采用PCR荧光分子信标技术检测YMDD耐药突变株 (包括YIDD YVDD突变株 )。结果表明 ,口服拉米夫定 1 2个月或 2 4个月 ,近半数病人血清HBVDNA转阴 (小于 0 1pg ml) ,35 % ( 2 7 77)的病人出现e抗原血清转换或单纯e抗原转阴 ,而e抗原血清转换绝大多数发生于HBVDNA低于 1 0 0pg ml血清的病人。 30 % ( 2 3 76)的病人出现YMDD耐药 (YIDD YVDD) ,在HBVDNA小于 1 0pg ml( 1 0 6拷贝 ml)的病人中YMDD耐药株与野生株共存。加大口服拉米夫定剂量 ,绝大多数YMDD耐药株仍不能转变为YMDD野生株。

To establish the method of molecular beacon and investigate the emergence and development of HBV resistant to lumivudine (YMDD resistant mutation). 77 hepatitis B patients were examined ALT, immune index, HBV DNA level and YMDD resistant mutation before and after treated with lumivudine orally for 12 months and 24 months. And YMDD resistant mutations, including YIDD and YVDD, were detected by the method of molecular beacon. After treated with lumivudine orally for 12 or 24 months, nearly half of the patients occurred seroconversion to HBV DNA level negative (<0 1pg/ml), 35% patients occurred HBeAg seroconversion (HBeAg negative and antiHBe positive) or only HBeAg negative without conversion to antiHBe positive, and most of these HBeAg seroconversions were occurred in patients with HBV DNA level lower than 100pg/ml. 30% patients occurred YMDD(YIDD/YVDD) resistant mutations, and YMDD resistant mutants coexisted with wild strains in patients with HBV DNA level lower than 10pg/ml. Most of the YMDD resistant mutants can not be converted to YMDD wild strains by increasing the dosage of lumivudine. It indicated that Chinese hepatitis patients were unfitted to increase dosage of lumivydine after occurred YMMD resistant mutation.