牛轮状病毒三种核酸检测方法的比较

Comparison of three nucleic acid amplification assays for bovine rotavirus

利用针对牛轮状病毒(BRV)的普通PCR、实时荧光定量PCR(Real-time PCR)和环介导等温扩增技术(LAMP)三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检测,比较三种核酸检测方法的检出结果。三种核酸检测方法中LAMP最敏感敏感,能检测到1拷贝/μL,Real-time PCR能检测到100拷贝/μL,普通PCR只能检测到1×104拷贝/μL。但结合临床样品检测表明:常规PCR方法敏感度低会造成一部分漏检,LAMP灵敏度高又会造成错检。综合比较三种方法后,推荐用LAMP结合real-time PCR,不仅节约成本,且结果更为准确可靠,可提高牛轮状病毒的检出率。

10-fold dilution series samples and 417 clinical samples were used in this study to evaluate the three BRV-specific detected methods： conventional PCR,real-time PCR and LAMP. The results showed that LAMP is the most sensitive method with its detection limit of 1 copies/μL,real-time PCR is 10copies/μL,but PCR had a lower sensitivity with its detection limit of 10000 copies/μL. Clinically,a contrast analysis of these three methods has learnt： the PCR had a higher undetected rate due to its low sensitivity to clinical samples;the error rate of LAMP was high due to its high sensitivity. These results suggested that the method of LAMP combining with real-time PCR is recommended in clinical detection of BRV,which is not only cost efficient but also accurate.