本文選題：豬流行性腹瀉 + 豬傳染性性胃腸炎��； 參考：《西北農(nóng)林科技大學》2017年碩士論文

【摘要】：豬流行性腹瀉病毒(porcine epidemic diarrhea virus,PEDV)和豬傳染性胃腸炎病毒(transmissible gastroenteritis virus,TGEV)是豬病毒性腹瀉的主要病原,不同年齡段的豬(尤其是仔豬)均易感,豬感染后以腹瀉、脫水、嘔吐為主要臨床特征,PEDV和TGEV已成為降低仔豬成活率的重要原因之一,對世界各國的養(yǎng)豬業(yè)造成了非常嚴重的經(jīng)濟損失。為了解陜西省PEDV和TGEV的流行病學及病原學特征,明確陜西省的流行毒株與疫苗株、經(jīng)典毒株和國內(nèi)外流行毒株的親緣關系及變異情況,進而為陜西省綜合防控PEDV和TGEV及研發(fā)更加合理有效的疫苗提供理論依據(jù)及參考數(shù)據(jù),本研究擬對陜西省部分地區(qū)進行PEDV和TGEV的流行病學調(diào)查,并對這兩種病毒的遺傳變異情況進行分析。研究取得如下結(jié)果。1.利用PCR法檢測了714份腹瀉病料中的PEDV。結(jié)果顯示,檢測出PEDV陽性病料288份,PEDV的陽性率為40.33%。在西安、咸陽、渭南、寶雞地區(qū)病料中均檢測到了PEDV,其中西安地區(qū)病料中PEDV陽性率最高,為46.94%。不同季節(jié)采集的病料中均檢測到了PEDV,其中冬季病料中PEDV陽性率最高,為57.96%。采集不同生長時期豬群的病料中也均檢測出了PEDV,其中哺乳仔豬的PEDV陽性率最高,為70.42%。2.利用PCR法檢測714份腹瀉病料中的TGEV。結(jié)果顯示,檢測出TGEV陽性病料69份,TGEV陽性率為9.97%,其中PEDV與TGEV混合感染的陽性病料20份,PEDV與TGEV混合感染陽性率為2.81%。在西安、咸陽、渭南、寶雞地區(qū)病料中均檢測到了TGEV,其中咸陽地區(qū)病料中TGEV的陽性率最高,為14.34%。不同季節(jié)采集的病料中均檢測出了TGEV,其中冬季病料中TGEV的陽性率最高,為17.05%。采集哺乳仔豬、斷奶仔豬、育肥豬、經(jīng)產(chǎn)母豬的病料中均檢測出了TGEV,陽性率分別為15.88%、8.52%、4.45%、7.29%,而后備母豬和種公豬中未檢測到TGEV。3.將處理好的PEDV陽性病料接種Vero細胞進行病毒的分離,通過細胞病變觀察、PCR、電鏡觀察等方法鑒定病毒。結(jié)果表明,該病毒能使細胞變圓、皺縮,繼而出現(xiàn)細胞破碎、脫落、拉絲等病變,經(jīng)PCR和電鏡觀察鑒定為PEDV,并命名為PEDV-SX株。以疫苗株CV777的基因組序列為模板,利用Primer 5.0設計了15對相互重疊的引物,進行全基因組的擴增及測序。利用DNA star軟件中的SeqMan軟件對序列進行拼接,并用MegAlign軟件對目的基因進行同源性的比較和遺傳進化分析。結(jié)果表明,PEDV-SX株與疫苗株的同源性為98%,與經(jīng)典毒株的同源性為97.8%,與國內(nèi)外流行株的同源性為97.2%~99.9%。PEDV-SX株與2012中國山東分離的SD-M株和2008年中國江蘇分離的JS2008株處在系統(tǒng)發(fā)育樹的同一個分支上。4.通過對TGEV-SX株的S、M、N、E基因進行分子克隆和測序,成功獲得了S、M、N、E基因序列。利用DNA star軟件中的MegAlign軟件對目的基因進行同源性比較和遺傳進化分析。結(jié)果表明,TGEV-SX株的S、M、N、E基因與8株參考毒株的同源性分別為98.0%~99.5%、98.1%~100%、97.9%~99.8%、96.8%~99.2%。TGEVSX株的S基因與SC-Y株、SHXB株、virulent Purdue株、Purdue P115株同屬一個大的分支,而TGEV-SX株的M、N、E基因與Miller M6株、Miller M60株、H16株、TS株同屬一個大的分支。本研究發(fā)現(xiàn)在陜西省部分地區(qū)豬群中均有PEDV與TGEV感染,PEDV的感染率較高,且主要感染哺乳仔豬,并存在二者混合感染。同源性分析及遺傳進化分析發(fā)現(xiàn),PEDV-SX株與2012中國山東分離的SD-M株和2008年中國江蘇分離的JS2008株的親緣關系較近,而與疫苗株CV777親緣關系較遠。同時也發(fā)現(xiàn)TGEV-SX株S基因與SC-Y株、SHXB株、virulent Purdue株、Purdue P115株親緣關系較近,而TGEVSX株的M、N、E基因與H16、TS株、Miller株親緣關系較近。
[Abstract]:Porcine epidemic diarrhea virus (porcine epidemic diarrhea virus, PEDV) and swine infectious gastroenteritis virus (transmissible gastroenteritis virus, TGEV) are the main pathogens of porcine viral diarrhea. Pigs of different ages (especially piglets) are susceptible to diarrhea, dehydration and vomiting as the main clinical characteristics after swine infection, PEDV and TGEV have become descending. One of the important reasons for the survival rate of low piglets has caused very serious economic loss to the pig industry of all countries in the world. To understand the epidemiological and pathogenic characteristics of PEDV and TGEV in Shaanxi Province, the relationship and variation of the epidemic strains and vaccine strains in Shaanxi Province, the classic strains and the epidemic strains at home and abroad, and the variation of the epidemic strains in Shaanxi province are also made. The prevention and control of PEDV and TGEV and the development of more reasonable and effective vaccines provide theoretical basis and reference data. This study intends to carry out an epidemiological survey of PEDV and TGEV in parts of Shaanxi Province, and analyze the genetic variation of these two viruses. The following results have been obtained by.1. and PCR method for detecting PEDV. in 714 diarrhoeal diseases. The results showed that 288 PEDV positive diseases were detected. The positive rate of PEDV was 40.33%. in Xi'an, Xianyang, Weinan and Baoji, and PEDV was detected in the disease materials in Baoji, among which the positive rate of PEDV was highest in Xi'an, and PEDV was detected in all the diseases collected in the different seasons of 46.94%.. The positive rate of PEDV in winter disease was the highest, which was 57.96%. mining. PEDV was also detected in the pigs of different growth periods, among which the positive rate of PEDV was the highest for suckling piglets. The TGEV. results in 714 diarrhoea samples were detected by 70.42%.2. using PCR method. The positive rate of TGEV positive was detected, the positive rate of TGEV was 9.97%, among which 20 were PEDV and TGEV mixed infection, PEDV and TGEV mixed feeling. The positive rate of 2.81%. was TGEV in Xi'an, Xianyang, Weinan and Baoji, among which the positive rate of TGEV was highest in Xianyang area, and TGEV was detected in all the diseases collected in different seasons. The positive rate of TGEV in winter disease was the highest, and 17.05%. collected suckling piglets, weanling piglets, fattening pigs, and production. TGEV was detected in the sows, and the positive rates were 15.88%, 8.52%, 4.45%, 7.29% respectively. In the back-up sow and the boar, the virus was isolated by the Vero cells that were treated with the PEDV positive disease, and the virus was identified by the pathological observation, PCR and electron microscopy. The results showed that the virus could make the cells change. The circle, crinkle, and then cell breakage, shedding, drawing and other lesions were identified as PEDV by PCR and electron microscopy, and named PEDV-SX strain. The genome sequence of the vaccine strain CV777 was used as a template, and Primer 5 was used to design 15 pairs of overlapping primers for whole genome expansion and sequencing. The sequence of SeqMan software in DNA Star software was used to sequence the sequence. The homologous comparison and genetic evolution analysis of the target genes were carried out with MegAlign software. The results showed that the homology of PEDV-SX strain and vaccine strain was 98%, the homology with the classic strains was 97.8%, and the homology of the domestic and foreign epidemic strains was 97.2%~99.9%.PEDV-SX strains isolated from 2012 China Shandong and Jiangsu of China in 2008. The isolated JS2008 strain was located on the same branch of the phylogenetic tree and.4. was sequenced by molecular cloning and sequencing of the S, M, N and E genes of the TGEV-SX strain. The sequence of S, M, N, E gene was successfully obtained. The homology of 8 strains of reference strain was 98.0%~99.5%, 98.1%~100%, 97.9%~99.8%, and 96.8%~99.2%.TGEVSX strain of S and SC-Y strain, SHXB strain, virulent Purdue strain and Purdue P115 strain. PEDV and TGEV infection were found among the pigs in some regions of western province, and the infection rate of PEDV was high, and the main infection was lactation piglets, and there were two mixed infections. Homology analysis and genetic evolution analysis found that the relationship between PEDV-SX strain and 2012 SD-M isolates from Shandong in China and JS2008 strains isolated from China in 2008 was close, and the vaccine strain CV77 was similar to the vaccine strain CV77. At the same time, it was found that the S gene of TGEV-SX strain was close to SC-Y, SHXB, virulent Purdue and Purdue P115, while M, N, and E genes were close.

【學位授予單位】：西北農(nóng)林科技大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S858.28

【相似文獻】

相關期刊論文 前10條

1 李凱年;用減毒PEDV防制仔豬PED[J];畜牧獸醫(yī)科技信息;2000年03期

2 余麗蕓;趙志剛;侯喜林;曹宏偉;王桂華;;乳酸菌表面表達PEDV S1蛋白的研究[J];黑龍江八一農(nóng)墾大學學報;2008年04期

3 高慎陽;劉延慶;李一經(jīng);;豬流行性腹瀉病毒(PEDV)M蛋白原核表達載體的構(gòu)建[J];中國獸醫(yī)學報;2009年12期

4 古洪浪;鄧勝朝;崔進;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

5 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)學報;2014年03期

6 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用[J];中國畜牧獸醫(yī);2014年01期

7 ;PEDV已出現(xiàn)了部分基因變異[J];畜禽業(yè);2014年01期

8 栗柱;;美國養(yǎng)豬業(yè)探討飼料在預防PEDV擴散中的作用[J];獸醫(yī)導刊;2014年06期

9 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];病毒學報;2009年03期

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

相關會議論文 前9條

1 余麗蕓;侯喜林;;Development and Evalutaion of Enzyme-Linked mmunosorbent Assay Based on Recombinant Nucleocapsid Protein for Detection of Porcine Epidemic Diarrhea (PEDV) Antibodies[A];中國畜牧獸醫(yī)學會家畜寄生蟲學分會第五次代表大會暨第八次學術研討會論文集[C];2004年

2 王金洛;宋維平;徐福洲;楊兵;賴平安;王琴;孟燕妮;;抗豬流行性腹瀉病毒卵黃抗體(PEDV-IgY)及其特性研究[A];中國畜牧獸醫(yī)學會2003年學術年會論文集[C];2003年

3 孫東波;馮力;時洪艷;陳建飛;;豬流行性腹瀉病毒分子生物學研究進展[A];中國畜牧獸醫(yī)學會畜牧獸醫(yī)生物技術學分會暨中國免疫學會獸醫(yī)免疫分會第七次研討會論文集[C];2008年

4 孫東波;朗洪武;時洪艷;陳建飛;崔曉辰;王承寶;佟有恩;馮力;;PEDV S蛋白B細胞抗原表位的篩選和鑒定[A];中國畜牧獸醫(yī)學會畜牧獸醫(yī)生物技術學分會暨中國免疫學會獸醫(yī)免疫分會第七次研討會論文集[C];2008年

5 鄭逢梅;趙軍;霍金耀;李歡;楊霞;常洪濤;杜季梅;王曉夢;王川慶;;PEDV流行株N基因主要抗原表位原核表達及ELISA方法的建立[A];中國畜牧獸醫(yī)學會家畜傳染病學分會第八屆全國會員代表大會暨第十五次學術研討會論文集[C];2013年

6 ;Identification of two novel B cell epitopes on porcine epidemic diarrhea virus spike protein[A];中國畜牧獸醫(yī)學會畜牧獸醫(yī)生物技術學分會暨中國免疫學會獸醫(yī)免疫分會第七次研討會論文集[C];2008年

7 李嘉愛;賴月輝;鄒永新;羅映霞;吳文福;巫月生;;PEDV免疫母雞血清抗體和蛋黃抗體的消長規(guī)律及其相關性測定[A];中國畜牧獸醫(yī)學會2003年學術年會論文集[C];2003年

8 ;Spike protein region (aa 636～789) of porcine epidemic diarrhea virus is essentIal for induction of neutralizing antibodies[A];中國畜牧獸醫(yī)學會畜牧獸醫(yī)生物技術學分會暨中國免疫學會獸醫(yī)免疫分會第七次研討會論文集[C];2008年

9 董世娟;朱于敏;于瑞嵩;李震;;我國PEDV分子流行病學研究進展[A];中國畜牧獸醫(yī)學會家畜傳染病學分會第八屆全國會員代表大會暨第十五次學術研討會論文集[C];2013年

相關博士學位論文 前10條

1 石達;宿主蛋白NPM1、EB3和HSP47對PEDV復制影響[D];中國農(nóng)業(yè)科學院;2015年

2 曹麗艷;豬流行性腹瀉病毒感染豬小腸上皮細胞抑制IFN-β產(chǎn)生及激活NF-κB機理研究[D];東北農(nóng)業(yè)大學;2015年

3 張月;豬流行性腹瀉病毒S基因變異分析及嗜酸乳桿菌口服疫苗的研究[D];山東農(nóng)業(yè)大學;2016年

4 李任峰;河南省豬流行性腹瀉病毒遺傳進化分析及免疫學檢測方法的建立[D];西北農(nóng)林科技大學;2016年

5 王大會;豬源益生菌對宿主腸道病毒的抗病毒作用研究[D];西北農(nóng)林科技大學;2016年

6 李R，

本文編號：1805175