【摘要】：為了解中國豬流行性腹瀉病毒(Porine epidemic diarrhea virus,PEDV)的流行現(xiàn)狀,本研究對上海和江蘇地區(qū)豬場進(jìn)行了PEDV流行株的分離鑒定。根據(jù)GenBank中PEDV的基因序列,對其M基因設(shè)計(jì)檢測引物。檢測臨床采集的豬腹瀉樣品,2份臨床樣品能擴(kuò)增出特異性目的條帶,測序結(jié)果證實(shí)為PEDV M基因片段。將檢測為PEDV陽性的樣品接種Vero細(xì)胞,并添加不同濃度的胰酶進(jìn)行病毒傳代培養(yǎng),在添加適量胰酶的Vero細(xì)胞上分離到兩株病毒并能穩(wěn)定傳代、增殖。選擇第22代病毒飼喂新生仔豬,進(jìn)行回歸試驗(yàn),結(jié)果兩株細(xì)胞毒均能使1日齡仔豬出現(xiàn)典型腹瀉癥狀,進(jìn)一步證實(shí)成功分離到PEDV,命名為JSLS/PEDV/1/2014和JS/PEDV/2/2014株。遺傳進(jìn)化分析,JS/PEDV/2/2014和JSLS/PEDV/1/2014分離株的M基因分別與中國毒株HLJ-2012、BJ-2012-1親緣性最近;S基因與I群PEDV流行毒株親緣性最近;兩個(gè)毒株的ORF3基因均存在51個(gè)堿基缺失,與DR13處于同一大分支,與CV777親緣性較遠(yuǎn)。
[Abstract]:In order to understand the epidemic status of porcine epidemic diarrhea virus (Porine epidemic diarrhea virus,PEDV) in China, PEDV epidemic strains were isolated and identified from pig farms in Shanghai and Jiangsu. According to the sequence of PEDV gene in GenBank, primers were designed to detect its M gene. The specific target bands were amplified from 2 clinical samples of porcine diarrhea, which were confirmed to be PEDV M gene fragments by sequencing. The PEDV positive samples were inoculated into Vero cells, and the virus was subcultured with different concentrations of trypsin. Two strains of viruses were isolated from Vero cells supplemented with proper amount of trypsin and could be subcultured and proliferated stably. The 22 ~ (th) generation virus was selected to feed newborn piglets. The results showed that both of the two strains could induce typical diarrhea symptoms in 1-day-old piglets. It was further confirmed that PEDV, was successfully isolated from piglets named JSLS/PEDV/1/2014 and JS/PEDV/2/2014 strains. Genetic evolution analysis showed that M gene of JS/PEDV/2/2014 and JSLS/PEDV/1/2014 isolate was most closely related to HLJ-2012,BJ-2012-1 of Chinese strain, S gene was most closely related to group I PEDV epidemic strain, and the ORF3 gene of both strains had 51 base deletion, which was at the same branch as DR13. The phylogenetic relationship with CV777 was far away.
【作者單位】： 上海市農(nóng)業(yè)科學(xué)院畜牧獸醫(yī)研究所;上海市農(nóng)業(yè)遺傳育種重點(diǎn)實(shí)驗(yàn)室;
【基金】：上海市科技興農(nóng)重點(diǎn)攻關(guān)項(xiàng)目(項(xiàng)目號(hào):滬農(nóng)科攻字(2013)第3-6號(hào));上海市科技興農(nóng)重點(diǎn)攻關(guān)項(xiàng)目(項(xiàng)目號(hào):滬農(nóng)科攻字(2013)第5-5號(hào)) 上海市生豬產(chǎn)業(yè)技術(shù)體系建設(shè)項(xiàng)目 上海市揚(yáng)帆計(jì)劃(項(xiàng)目號(hào):15YF1410300) 上海市農(nóng)口青年成長計(jì)劃項(xiàng)目(項(xiàng)目號(hào):滬農(nóng)青字(2016)第1-35號(hào))
【分類號(hào)】：S852.65

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 古洪浪;鄧勝朝;崔進(jìn);劉曉露;劉超;袁航;馬嵐;張桂紅;;豬流行性腹瀉病毒PEDV-GD12株的分離與鑒定[J];中國獸醫(yī)雜志;2013年11期

2 杜曉莉;王一成;吳潤;徐麗華;袁秀芳;李軍星;周曉麗;;2010—2013年浙江省豬流行性腹瀉病毒臨床檢測及PEDV-S基因型分析[J];浙江農(nóng)業(yè)學(xué)報(bào);2014年03期

3 鄧祖麗穎;陳陸;;豬流行性腹瀉病毒巢式RT-PCR檢測方法的建立及應(yīng)用[J];中國畜牧獸醫(yī);2014年01期

4 李寶賢;馬廣鵬;葛俊偉;李一經(jīng);;豬流行性腹瀉病毒功能性受體的鑒定[J];病毒學(xué)報(bào);2009年03期

5 ;Cloning the Structure Genes and Expression the N Gene of Porcine Epidemic Diarrhea Virus DX[J];Virologica Sinica;2009年03期

6 喬憲鳳,熊忠良,華文君,鄭新民;PEDV膜蛋白基因RT-PCR最適反應(yīng)條件的確立[J];湖北畜牧獸醫(yī);2001年01期

7 高慎陽;王s，

本文編號(hào)：2264606