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猪德尔塔冠状病毒纳米PCR检测方法的建立

点击数:7605  时间:2019-08-13 16:56:40  来源: 信息处      

根据GenBank登录的猪德尔塔冠状病毒(PDCoV)毒株M基因设计特异性引物,建立了PDCoV纳米PCR检测方法并进行了临床样品检测。结果显示:该方法对标准品检测的最低浓度可达102 copies/μL,与其他几种常见猪病病毒无交叉反应;用该方法检测国内猪场62份腹泻粪便样品,相比常规PCR方法,该纳米PCR方法的检出率更高,而两种方法对268份进境活猪粪拭子样品检测均为阴性。结果表明,本研究所建立的纳米PCR检测方法适用于国内猪场的PDCoV检测和进境活猪的监控检测,可作为PDCoV实验室检测的有力技术手段。

Establishment of Nanoparticle-assisted PCR Assay for Detection ofPorcine Deltacoronavirus

In this paper,a pair ofprimers were designed according to the M gene sequence of porcinedeltacoronavirus(PDCoV)that waspublished in Genbank,then a kind ofnanoparticle-assisted PCR assay was established. The results showed that thestandard samples with minimum concentration of 102 copies/μL could be detected by this method,and therewas no any cross-reaction with other common swine disease viruses. Compared toregular PCR assay,the nanoparticle-assisted PCR assaypresented a higher detection rate when 62 diarrhea feces collected fromdomestic swine farms were tested. When 268 fecal swab samples of imported livepigs were tested by both the two methods,the detectionresults all showed negative. Therefore,thenanoparticle-assisted PCR assay established in the paper could be used indetecting PDCoV in domestic swine farms and monitoring imported live pigs,which would be a strong technical tool for laboratory detection ofPDCoV.

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