猪丁型冠状病毒荧光定量RT-PCR与S1蛋白间接ELISA检测方法的建立及应用

发布时间：2018-04-25 07:36

本文选题：猪丁型冠状病毒 + 实时荧光定量RT-PCR　；参考：《生物工程学报》2017年08期

【摘要】：最近,新发现的猪丁型冠状病毒(Porcine deltacoronavirus,PDCo V)可导致仔猪腹泻,亟待建立有效准确的核酸及血清学检测方法并调查猪场的PDCo V感染情况。本研究采用针对病毒M基因的探针荧光定量RT-PCR检测方法,对2014-2015年间收集的河南、湖南、浙江、江西、安徽、河北、黑龙江、江苏、山东和上海等10个省市收集的共254份腹泻仔猪小肠匀浆或粪便样本进行检测,共检出11份PDCo V阳性样本,总阳性率为4.33%。采用在昆虫细胞表达纯化的PDCo V囊膜S1蛋白为包被抗原,建立间接酶联免疫吸附试验(ELISA)方法,检测2015-2016年间收集的来自江苏、湖南、浙江等7个省份的609份具有腹泻症状的猪血清样品,抗PDCo V S1抗体检出率为44.17%(269/609)。上述建立的两种方法分别针对病毒的RNA或抗体对PDCo V进行特异性检测,可以应用于PDCo V流行情况的监控。
[Abstract]:Recently, porcine deltacoronavirusPDCo V, a newly discovered porcine coronavirus, can cause diarrhea in piglets. It is urgent to establish an effective method for nucleic acid and serological detection and to investigate the PDCo V infection in pig farms. In this study, fluorescence quantitative RT-PCR was used to detect virus M gene in Henan, Hunan, Zhejiang, Jiangxi, Anhui, Hebei, Heilongjiang and Jiangsu provinces during 2014-2015. A total of 254 intestinal homogenate or fecal samples of diarrhea piglets were collected from 10 provinces and cities, including Shandong and Shanghai, and 11 PDCo V positive samples were detected. The total positive rate was 4.33. An indirect enzyme-linked immunosorbent assay (Elisa) was established to detect the expression of PDCo V envelope S1 protein in insect cells from Jiangsu, Hunan, and Hunan during the period of 2015-2016, and to establish an indirect enzyme-linked immunosorbent assay (Elisa) method. The detection rate of anti PDCo V S1 antibody in 609 porcine serum samples with diarrhea symptoms in 7 provinces of Zhejiang Province was 44.17 / 269 / 609. The above two methods can be used to monitor the prevalence of PDCo V by specific detection of PDCo V against RNA or antibody.
【作者单位】：浙江大学动物科学学院动物预防医学研究所;杭州贝尔塔生物技术有限公司;
【基金】：国家重点研究发展计划(No.2016YFD0500102)资助~~
【分类号】：S852.651

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