新疆南疆一起PPR的診斷及PPRV基因序列分析
發(fā)布時(shí)間:2018-03-05 02:34
本文選題:小反芻獸疫 切入點(diǎn):診斷 出處:《塔里木大學(xué)》2017年碩士論文 論文類型:學(xué)位論文
【摘要】:小反芻獸疫(Peste des Petits Ruminants,PPR)是由副黏病毒科(Paramyxoviridae)麻疹病毒屬的小反芻獸疫病毒(Peste des Petits Ruminants Virus,PPRV)引起的急性、烈性、高度接觸性傳染病,臨床表現(xiàn)為發(fā)熱、呼吸困難、口鼻分泌物增多、水樣腹瀉等。2014年5月新疆南疆阿克蘇地區(qū)某羊場(chǎng),先后有多只羊出現(xiàn)高熱、精神沉郁、口鼻大量灰白色惡臭狀分泌物、呼吸困難、拉稀、消瘦等癥狀。通過(guò)對(duì)病羊的臨床診斷、病理學(xué)檢查,初步懷疑該群羊發(fā)生PPRV感染。通過(guò)福爾馬林固定肺臟組織中核酸的提取及PPRV N基因序列的擴(kuò)增,并對(duì)PCR產(chǎn)物測(cè)序結(jié)果進(jìn)行Blast比對(duì)。結(jié)果顯示,該基因片段與中國(guó)第二次PPR疫情流行毒株的同源性高達(dá)99%,綜合確診為該羊場(chǎng)受到PPRV感染。實(shí)驗(yàn)室對(duì)福爾馬林固定肺臟組織進(jìn)行核酸提取及PPRV全基因序列擴(kuò)增,獲得新疆南疆PPRV毒株全基因序列并命名為China XJNJ2014。對(duì)該毒株進(jìn)行全基因及N、F基因序列分析,全基因序列分析顯示:本試驗(yàn)毒株與國(guó)內(nèi)吉林GZL-14毒株同源性最高,與國(guó)外印度Izatnagar/94毒株及Sungri1996 MSD毒株同源性最高。N基因核苷酸序列分析顯示:試驗(yàn)毒株與中國(guó)新疆China/XJYL/2013毒株及中國(guó)廣東GD/QY/2014毒株進(jìn)化關(guān)系最近,與國(guó)外印度IND/Delhi/2016/05毒株進(jìn)化關(guān)系最近。F基因核苷酸序列分析顯示:試驗(yàn)毒株與國(guó)內(nèi)北京China/BJ/2014毒株、吉林GZL-14毒株和浙江PPRV-FY毒株進(jìn)化關(guān)系最近,與國(guó)外印度Izatnagar/94毒株和Izatnagar-94毒株進(jìn)化關(guān)系最近。全基因及N、F基因核苷酸分析均顯示試驗(yàn)毒株與西藏毒株及中國(guó)目前使用的疫苗毒株進(jìn)化關(guān)系較遠(yuǎn)。N蛋白分析顯示:試驗(yàn)毒株與中國(guó)河南CH/HNNY/2014毒株、CH/HNZM/2014毒株、中國(guó)廣東GD/QY/2014毒株、中國(guó)浙江PPRV-FY毒株及國(guó)外的印度IND/Delhi/2016/05毒株進(jìn)化關(guān)系最近。F蛋白氨基酸序列分析顯示:試驗(yàn)毒株與河南CH/HNZK/2014毒株、北京China/BJ/2014毒株、吉林GZL-14毒株和浙江PPRV-FY毒株及國(guó)外印度Izatnagar/94毒株進(jìn)化關(guān)系最近。N、F蛋白分析顯示試驗(yàn)毒株與西藏毒株及中國(guó)目前使用的疫苗毒株進(jìn)化關(guān)系較遠(yuǎn)。全基因及N、F基因核苷酸進(jìn)化樹(shù)構(gòu)建結(jié)果均顯示試驗(yàn)毒株與中國(guó)第二次疫情流行毒株同屬第Ⅳ系的一個(gè)小分支,與國(guó)外的印度毒株進(jìn)化關(guān)系較近。通過(guò)本研究表明,新疆阿克蘇地區(qū)的本次疫情為PPRV感染綿羊所致,通過(guò)基因序列分析表明本試驗(yàn)毒株與中國(guó)2013~2014年P(guān)PR大流行時(shí)期的毒株進(jìn)化關(guān)系最高,系統(tǒng)進(jìn)化關(guān)系顯示此次疫情毒株與中國(guó)2013~2014年P(guān)PR大流行時(shí)期的毒株同處于一個(gè)分支(第Ⅳ譜系)。本研究結(jié)果不僅能為該地區(qū)PPR診斷提供參考,而且能有效的為該地區(qū)的疫情防控提供理論依據(jù)。
[Abstract]:Peste des Petits Ruminants (PPRV) is an acute, acute and highly contagious disease caused by the measles virus of the genus Paramyxoviridae, Peste des Petits Ruminants VirusPPRV.Clinical manifestations include fever, dyspnea, and increased oral and nasal secretions. In May 2014, a certain sheep farm in Aksu area, southern Xinjiang, had several sheep with high fever, depressed spirit, a large amount of greyish-white stinking secretion, dyspnea, dilation, wasting and other symptoms. Through the clinical diagnosis of the sick sheep, Pathological examination showed that the sheep were suspected to be infected with PPRV. The nucleic acid extracted from formalin fixed lung tissue and the amplification of PPRV N gene sequence were used to compare the PCR product sequencing results with Blast. The homology of the gene fragment with the second epidemic strain of PPR in China was as high as 99%. It was confirmed that the sheep farm had been infected with PPRV. The nucleic acid extracted from formalin fixed lung tissue and the whole PPRV gene sequence were amplified in laboratory. The whole gene sequence of PPRV strain in southern Xinjiang was obtained and named China XJNJ2014.The whole gene sequence analysis of this strain showed that this strain had the highest homology with Jilin GZL-14 strain in China. The nucleotide sequence analysis of the highest homology of .N gene with Indian Izatnagar/94 strain and Sungri1996 MSD strain showed that the experimental strain had the closest evolutionary relationship with Xinjiang China/XJYL/2013 strain in China and GD/QY/2014 strain in Guangdong Province, China. The nucleotide sequence analysis of the. F gene showed that the experimental strain had the closest evolutionary relationship with the domestic Beijing China/BJ/2014 strain, Jilin GZL-14 strain and Zhejiang PPRV-FY strain. The nucleotide analysis of the whole gene and NV F gene showed that the phylogenetic relationship between the tested strain and the Tibetan strain and the vaccine strain currently used in China was far away from that of the foreign strains of Indian Izatnagar/94 and Izatnagar-94. The results of nucleotide analysis of the whole gene and the NV F gene showed that there was a distant relationship between the tested strains and the vaccine strains used in China. The virus test strain and the Henan CH/HNNY/2014 virus strain CHP HNZM / 2014 strain, Amino acid sequence analysis of the most recent. F protein amino acid sequences of Guangdong GD/QY/2014 strain, Zhejiang PPRV-FY strain in China and Indian IND/Delhi/2016/05 strain in foreign countries showed that the tested strain was associated with Henan CH/HNZK/2014 strain and Beijing China/BJ/2014 strain. The evolutionary relationship between Jilin GZL-14 strain, Zhejiang PPRV-FY strain and foreign Indian Izatnagar/94 strain. The analysis of the protein of the tested strain shows that the relationship between the tested strain and the Tibetan strain and the vaccine strain currently used in China is far away. The nucleotides of the whole gene and the NNF gene are far away from that of the Tibetan strain and the vaccine strain currently used in China. The results of phylogenetic tree construction showed that the tested strain was a small branch of the fourth strain of the second epidemic in China. This study showed that the epidemic situation in Aksu region of Xinjiang was caused by PPRV infection in sheep. The genetic sequence analysis showed that this strain had the highest evolutionary relationship with the strain in the period of PPR pandemic from 2013 to 2014 in China. The phylogenetic relationship showed that the epidemic strain was in the same branch as the strain in the period of PPR epidemic from 2013 to 2014 in China. The results of this study can not only provide reference for the diagnosis of PPR in this area, but also for the diagnosis of PPR in this region. And can effectively provide the theoretical basis for the prevention and control of the epidemic situation in this area.
【學(xué)位授予單位】:塔里木大學(xué)
【學(xué)位級(jí)別】:碩士
【學(xué)位授予年份】:2017
【分類號(hào)】:S858.26
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1 張路瑤;新疆南疆一起PPR的診斷及PPRV基因序列分析[D];塔里木大學(xué);2017年
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