牛病毒性腹泻病毒三种核酸检测方法的比较

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。

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

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.