乙型肝炎病毒YMDD变异不同检测方法比较与评价  

Comparison and evaluation study on detection of YMDD mutation in hepatitis B virus with different methods

作　　者：李红梅[1] 彭忠田[1] 刘书香[1] 邱红梅[1] 

机构地区：[1]南华大学附属第一医院肝病研究中心,湖南衡阳421001

出　　处：《中国现代医学杂志》2009年第23期3532-3536,共5页China Journal of Modern Medicine

摘　　要：目的比较检测乙型肝炎病毒(HBV)YMDD变异的3种不同方法,评价其在监测拉米夫定抗HBV治疗中发生耐药突变的价值。方法对80例接受拉米夫定治疗的慢性乙型肝炎患者,测定其治疗前后血清HBVDNA含量、ALT水平变化,分别用聚合酶链反应微板核酸杂交-酶联免疫吸附法(PCR-ELISA)、通用模板信号扩增(UT-PCR)技术、以及基因测序3种方法进行HBVYMDD变异检测,并分析结果。结果80例患者用基因测序法检测出34例发生YMDD变异,变异发生率为41.25%;用PCR-ELISA法检测出21例变异阳性,与基因测序检测出的阳性率差异有显著性(χ2=4.68,P<0.05),两者结果的符合率为61.76%;用UT-PCR法检测出33例变异阳性,与基因测序检测出的阳性率无统计学差异(χ2=0.03,P>0.05),两者结果的符合率为97.06%。结论基因测序和UT-PCR方法检测HBVYMDD变异非常可靠,是监测拉米夫定耐药株的非常有效的方法,UT-PCR方法更适合临床应用,而PCR-ELISA方法需提高其敏感性。【Objectives】To compare different methods that detected the YMDD mutation in hepatitis B virus （HBV）, and to evaluate them in monitoring the Lamivudine-resistant HBV mutants. 【Methods】In 80 patients with chronic hepatitis B, the level of HBV DNA and ALT were detected before and after Lamivudine treatment. HBV YMDD mutation was detected and analysed by 3 different methods： PCR-ELISA, UT-PCR, and DNA sequence. 【Results】34 YMDD mutation were deteced by the means of DNA sequence, the mutation occurrence rate is 41.25%. 21 positive cases were detected by the means of PCR-ELISA, there are significant difference in the positive rate eompare with DNA sequenee （X^2=4.68, P 〈0.05）, the rate of consistency these two method was 61.75%. 33 positive eases were detected by the means of UT-PCR, there are no significant differenee in the positive rate eompare with DNA sequence （X^2=0.03, P 〉0.05）, the rate of consistency these two method was 97.06%. [ Cochtsions] The DNA sequence and UT-PCR assay were reliable to detect HBV YMDD mutations, and they were effeetive in monitoring the Lamivudine-resistant HBV mutants. UT-PCR is more applicable to clinical laboratory use. PCR-ELISA should be further improved to increase sensitivity.

关 键 词：肝炎病毒 乙型 YMDD变异 聚合酶链反应微板核酸杂交-酶联免疫吸附 通用模板信号扩增 基因测序 

分 类 号：R446.5[医药卫生—诊断学]