【摘要】：牛病毒性腹瀉/粘膜病毒(Bovine viral diarrhea/mucosal disease virus,BVD/MDV)是黃病毒科,瘟病毒屬的成員,具有極其嚴(yán)重的危害性�？梢鹨环N呈世界性分布的疾病—牛病毒性腹瀉/粘膜病,給畜牧業(yè)造成了極大的危害。隨著畜牧業(yè)的快速發(fā)展,其危害性也日益嚴(yán)重。目前,該病尚未有有效的防控措施。因此,獲得牛病毒性腹瀉的流行病學(xué)資料,為該病的后續(xù)防控提供充足的理論基礎(chǔ)。本研究采用血清學(xué)方法和分子生物學(xué)方法對(duì)新疆16個(gè)地區(qū)不同牛場(chǎng)采集的血清、全血和糞便樣品進(jìn)行檢測(cè),并對(duì)檢測(cè)到的陽(yáng)性樣品進(jìn)行病原學(xué)分離鑒定。通過(guò)對(duì)1171份樣品進(jìn)行血清學(xué)檢測(cè),其中抗體陽(yáng)性率為81.9%;抗原陽(yáng)性率為1.5%。應(yīng)用RT-PCR方法對(duì)566份樣品進(jìn)行檢測(cè),從中檢測(cè)到了21株陽(yáng)性BVDV樣品。對(duì)測(cè)序結(jié)果進(jìn)行序列同源性比對(duì)分析,繪制其進(jìn)化樹(shù),并初步繪制出了新疆部分地區(qū)BVDV病毒的地理生態(tài)分布圖,其中17株為BVDV I型,4株為BVDV II型。應(yīng)用細(xì)胞培養(yǎng)分離病毒,利用RT-PCR方法和直接熒光免疫檢測(cè)進(jìn)一步鑒定病毒,最后測(cè)定病毒的病毒滴度和對(duì)陽(yáng)性血清的中和效價(jià),并進(jìn)行病毒的理化特性和生物學(xué)特性分析。結(jié)果發(fā)現(xiàn)7株為致細(xì)胞病變型,14株為非致細(xì)胞病變型。所測(cè)得的病毒滴度為1.04×103~6.02×108 TCID50/0.1mL,21株陽(yáng)性樣品對(duì)BVDV陽(yáng)性血清的中和效價(jià)為1:21~1:25。分離的病毒對(duì)溫度、乙醚、氯仿和胰酶很敏感。新疆地區(qū)的BVDV流行株有BVDV 1b亞型、I型未知亞型和BVDV II型。研究成果為BVDV的預(yù)防和控制提供了理論基礎(chǔ)和參考依據(jù)。
[Abstract]:Bovine viral diarrhea / mucosal virus (Bovine viral diarrhea/mucosal disease virus,BVD/MDV) is a member of the family Flavoviridae. It can cause a worldwide disease-bovine viral diarrhea / mucosal disease, which has caused great harm to animal husbandry. With the rapid development of animal husbandry, its harmfulness is becoming more and more serious. At present, there is no effective prevention and control measures. Therefore, to obtain the epidemiological data of bovine viral diarrhea provides sufficient theoretical basis for the follow-up prevention and control of the disease. In this study, serological and molecular biological methods were used to detect serum, whole blood and fecal samples collected from 16 cattle farms in Xinjiang, and the positive samples were isolated and identified by etiology. By serological examination of 1171 samples, the positive rate of antibody and antigen were 81.9 and 1.5 respectively. RT-PCR method was used to detect 566 samples, from which 21 positive BVDV samples were detected. The sequence homology analysis was carried out, the evolutionary tree was drawn, and the geographic and ecological distribution of BVDV virus in some regions of Xinjiang was preliminarily plotted. Among them, 17 strains were BVDV I type and 4 were BVDV II type. The virus was isolated by cell culture. The virus was further identified by RT-PCR method and direct fluorescence immunoassay. Finally, the virus titer and neutralization titer to positive serum were determined, and the physicochemical and biological characteristics of the virus were analyzed. The results showed that 7 strains were cytopathic and 14 were non-cytopathic. The titer of the virus was 1.04 脳 10 ~ 3 and 6.02 脳 10 ~ 8 TCID50/0.1mL,21 strain. The neutralization titer of BVDV positive serum was 1: 21 1: 25. The isolated virus is sensitive to temperature, ether, chloroform and trypsin. The research results provide theoretical basis and reference for prevention and control of BVDV.
【學(xué)位授予單位】：新疆農(nóng)業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：S858.23

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