牛病毒性腹泻病毒三种核酸检测方法的比较

Comparative study of three nucleic acid amplification assays for the detection of bovine viral diarrhea virus

利用针对牛病毒性腹泻病毒(BVDV)的常规RT-PCR、实时荧光定量RT-PCR(real-time RT-PCR)和RT-环介导等温扩增技术(RT-LAMP)3种核酸检测方法对不同质粒浓度的样品和417份临床疑似样品进行了检测,以比较3种核酸检测方法的优越性。结果显示,3种核酸检测方法中,real-time RT-PCR和RT-LAMP一样敏感,均能检测到10拷贝/μL,常规RT-PCR只能检测到1×104拷贝/μL,但临床样品检测表明,常规RT-PCR方法敏感度低,会造成一部分漏检,RT-LAMP灵敏度高又会造成错检。综合比较3种方法后,推荐用RT-LAMP结合real-time RT-PCR,不仅节约成本,且结果更为准确可靠,可提高牛病毒性腹泻的检出率。

Ten fold dilution series samples and 417 clinical samples suspected with bovine viral diarrhea were used to evaluate three BVDV（bovine viral diarrhea virus）-specific detection methods：conventional RT-PCR,real-time RT-PCR and RT-LAMP（loop-mediated isothermal amplification）.Results showed that the real-time RT-PCR and RT-LAMP had the same high sensitivity,and both of them could detect as lower as 10 copies/μL of samples,but the conventional RT-PCR had a lower sensitivity with its detection limit of 1×104 copies/μL.Clinically,a contrast analysis of these three methods showed that the RT-PCR had a higher undetected rate duo to its low sensitivity to clinical samples;the error rate of RT-LAMP was high due to its high sensitivity.These results suggested that the method of RT-LAMP combining with real-time RT-PCR is the recommended methods in clinical detection of BVDV,which is not only cost efficient but also accurate.