检测羊肠道病毒RT-PCR方法的建立及初步应用

Establishment and preliminary application of RT-PCR method for the detection of caprine/ovine enterovirus

羊肠道病毒(caprine enterovirus,CEV)感染为国内外新发疫病,其诊断方法尚缺乏研究。本研究依据CEV-JL14毒株的基因组序列,设计合成了1对特异性引物,建立了检测CEV的RT-PCR方法,并对其特异性、敏感性和重复性进行测定,同时应用该方法对疑似CEV感染的临床样品进行检测。结果显示,建立的检测CEV感染的RT-PCR方法具有特异性好、敏感性高、快速和简便等特点,最低检出限为1.13×10^(3)copies/μL。以建立的RT-PCR方法对通过ELISA方法确定的6份阳性样品和22份阴性样品进行检测,结果显示两者的符合率为100%。本研究建立的RT-PCR方法可用于CEV感染的诊断和流行病学调查。

Caprine enterovirus(CEV)infection is an emerging disease at home and abroad that causes a significant economic loss to goat/sheep industry.Currently,few methods are available for diagnosis of this infection.In this study,a pair of goat enterovirus-specific primers was designed and synthesized based on the genome sequence of goat enterovirus CEV-JL14 strain.RT-PCR method for the detection of goat enterovirus was established,and the specificity,sensitivity and repeatability were tested.In addition,the established RT-PCR methods was compared with a sandwich ELISA for the detection of clinical samples suspected of CEV infection.The results showed that the established RT-PCR method was specific for detecting CEV with a detection sensitivity of 1.13×10^(3)copies/μL.The established RT-PCR method was used to detect six positive samples and 22 negative samples determined by ELISA method,and the coincidence rate of the two methods was 100%.The above results indicate that the established RT-PCR method can be used for the diagnosis and epidemiological investigation of CEV infection.