六种检测猪瘟病毒方法的比较

Comparison of six detection methods for classical swine fever virus

摘要:

摘要：【目的】本研究旨在比较6种检测猪瘟病毒方法的优缺点。【方法】应用病毒分离、胶体金免疫层析试纸条、抗原捕捉ELISA、反转录-聚合酶链式反应（RT-PCR）、TaqMan荧光定量RT-PCR（RT-qPCR）和反转录-环介导等温扩增方法（RT-LAMP）等6种方法，分别对50份疑似猪瘟病料中的猪瘟病毒（Classical swine fever virus，CSFV）进行检测。【结果】结果表明：RT-qPCR和RT-LAMP方法检出阳性样品数为13份，RT-PCR为11份，病毒分离为10份，抗原捕捉ELISA为9份，胶体金试纸条为8份；6种方法均检测为阳性8份，均为阴性37份。【结论】结果提示，在对猪瘟病毒进行检测时，RT-qPCR、RT-LAMP和RT-PCR由于其灵敏性高，可作为首选检测方法，但操作时需要避免假阳性的出现；病毒分离方法虽然操作繁琐，但结果准确，是确诊猪瘟必不可少的检测方法；抗原捕捉ELISA和胶体金试纸条检测时间较短，由于其敏感性较低所限，主要用于对畜群进行检测，不适合个体检测。

Abstract:

Abstract: [Objective] We investigated the advantages and disadvantages of six methods to detect classical swine fever virus (CSFV). [Methods] We used six methods, including the virus isolation, colloidal gold immunochromatographic assay (CGIA), antigen-capture ELISA (AC-ELISA), reverse transcription?polymerase chain reaction (RT-PCR), TaqMan real-time RT-PCR (RT-qPCR) and reverse transcription?loop-mediated isothermal amplification assay (RT-LAMP), to detect CSFV in 50 samples parallelly. [Results] The results showed that 13 samples were detected positive by RT-qPCR and RT-LAMP, 11 by PCR, 10 by virus isolation, 9 by AC-ELISA and 8 by CGIA, and 8 samples were detected positive and 37 samples negative by the six methods. [Conclusion] These results indicated that the 3 RNA-amplification assays could be used as the first choice for detection of CSFV due to the high sensitivity, while they were vulnerable to false positive results arising from sample to sample contaminations or from other contaminated sources. Although the virus isolation was time-consuming, it was still considered the “gold standard” and was indispensable for confirming CSF outbreaks. The rest two methods, AC-ELISA and CGIA, yielded the results in a short time yet their performance was hampered by a low sensitivity. Therefore, they were mainly used for herd diagnosis and not suitable for individual test for CSFV infection.