逆转录环介导等温扩增检测风疹病毒RNA方法的建立

Establishmentof reverse-transcription-loop-mediated isothermal amplifi-cation method for detection of rubella virus

目的建立逆转录环介导等温扩增快速检测风疹病毒RNA的方法。方法选取保守基因E1设计引物,建立并优化RT-LAMP反应体系,同时进行特异性和敏感性评估。在此基础上应用建立的方法对46例ELISA检测阳性的孕妇血清标本进行检测,并与RT-PCR相比较。结果建立的方法仅对风疹病毒RNA扩增出特有的梯状条带,酶切结果与预期相符。46例孕妇标本经RT-PCR和RT-LAMP检测,分别检出28例和29例。结论该方法特异性强、敏感性高、简便快速且成本低,有望在临床检测中发挥一定的作用。

Objective: To establish a method of Reverse-transcription-loop-mediated isothermal Amplification(RT-LAMP) for detection of Rubella Virus RNA.Methods: Primers were designed according to the Rubella′s selfish gene E1,and then the system of RT-LAMP to detect Rubella Virus RNA were established and optimized.Meanwhile,the specificity and sensitivity were evaluated.Been identified by ELISA,the 46 positive blood serum samples from pregnant women were tested using this method,and the results were compared by RT-PCR.Results: The products of RT-LAMP demonstrated the typical ladder patterns only if the temple was Rubella Virus RNA,and digested fragments were identical with the predicted sizes.Among the 46 samples from pregnant women,the positive results turned out to be 28 and 29 by using RT-PCR and RT-LAMP respectively.Conclusions: The results indicated that this RT-LAMP system not only had good specificity,sensitivity,but simple,rapid and low-cost,and might play a role in clinical detection.