摘要
目的 建立一种在拉米夫啶治疗过程中的聚合酶酪氨酸 蛋氨酸 天冬氨酸 天冬氨酸(YMDD)基序变异的检测方法。方法 根据寡核苷酸探针与模板结合时 ,不匹配碱基显著影响熔解温度的原理 ,设计合成 2条荧光标记的特异性探针 ,用实时荧光聚合酶链反应 (PCR)方法对 2 97例患者的 35 4份标本进行YMDD变异检测。结果 35 4份标本中 ,检出YMDD变异株 15 6份 ( 44 1% ) ,其中混合株占 34% ( 5 3/15 6 ) ;有 9份血清标本同时进行了DNA序列分析 ,测序结果同本试验检测的结果完全一致 ;同时发现YMDD变异与血清谷丙氨酸转氨酶异常相关 (P <0 0 1)。结论 本研究建立的实时荧光PCR方法简便、快速、灵敏、经济 ,可用于临床拉米夫啶治疗中YMDD变异的检测。
Objective Setting up a method to detect YMDD mutation during lamivudine therapy. Methods Two specific fluorescent probes were designed, on the unpaired base can remarkably affect melting temperature when oligonucleotide probe binding to DNA template. 297 patients (354 sera samples) were detected YMDD mutation by real time fluorimetry PCR. Results 156 YMDD mutation were found in 354 sera samples (44.1%), in which the mixed variant species were 34%(53/156). In the same time, nine sera samples wre sequenced. The results of sequencing were fully corresponded to those of fluorimetry PCR. The statistical analysis showed that YMDD variants were obviously correlated with ALT level(P<0.01). Conclusion The method described here provides a convenient, rapid, accurate and inexpensive way to monitor YMDD mutation during lamivudine therapy.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2003年第1期37-39,共3页
Chinese Journal of Laboratory Medicine
