实时荧光定量PCR技术快速检测麻疹病毒核酸的研究

TaqMan-based Real-time PCR Assay for Quick Detection of Measles Virus RNA

目的 建立以特异性荧光探针为特点的TaqMan荧光定量RT PCR方法 ,对麻疹病毒的核酸进行检测。方法 对设计的引物与探针进行了筛选与条件优化 ,克服了常规RT PCR定性检测的不足。结果 具有对麻疹病毒检测的高度特异性与准确性 ,检测的敏感度可达 0 1TCID50 ,从病毒核酸提取至检测完成仅需 3个多小时 ,操作简便 ,而且大大减少了常规PCR扩增产物的污染机会。结论 采用该方法对麻疹病毒与相关的临床样本进行了检测 ,均获得了满意的结果 ,为麻疹病毒的分子生物学检测 。

Objective To establish a real time PCR assay based on Taq Man chemistry for detection of measles virus RNA. Methods The primers and probes were screened, and reactive condition was optimized. Results The specificity for detection of measles virus RNA was high, and the sensitivity was 0 1 TCID 50 . It only took three hours to complete the whole course, while the chance contamination decreased obviously compared to conventional PCR. Some clinic samples were detected by this method, and the results were satisfactory. Conclusions The TaqMen based real time PCR is rapid,sensitive and reliable for detection of measles virus.