新疆南疆一起PPR的诊断及PPRV基因序列分析

发布时间：2018-03-05 02:34

本文选题：小反刍兽疫　切入点：诊断　出处：《塔里木大学》2017年硕士论文　论文类型：学位论文

【摘要】：小反刍兽疫(Peste des Petits Ruminants,PPR)是由副黏病毒科(Paramyxoviridae)麻疹病毒属的小反刍兽疫病毒(Peste des Petits Ruminants Virus,PPRV)引起的急性、烈性、高度接触性传染病,临床表现为发热、呼吸困难、口鼻分泌物增多、水样腹泻等。2014年5月新疆南疆阿克苏地区某羊场,先后有多只羊出现高热、精神沉郁、口鼻大量灰白色恶臭状分泌物、呼吸困难、拉稀、消瘦等症状。通过对病羊的临床诊断、病理学检查,初步怀疑该群羊发生PPRV感染。通过福尔马林固定肺脏组织中核酸的提取及PPRV N基因序列的扩增,并对PCR产物测序结果进行Blast比对。结果显示,该基因片段与中国第二次PPR疫情流行毒株的同源性高达99%,综合确诊为该羊场受到PPRV感染。实验室对福尔马林固定肺脏组织进行核酸提取及PPRV全基因序列扩增,获得新疆南疆PPRV毒株全基因序列并命名为China XJNJ2014。对该毒株进行全基因及N、F基因序列分析,全基因序列分析显示:本试验毒株与国内吉林GZL-14毒株同源性最高,与国外印度Izatnagar/94毒株及Sungri1996 MSD毒株同源性最高。N基因核苷酸序列分析显示:试验毒株与中国新疆China/XJYL/2013毒株及中国广东GD/QY/2014毒株进化关系最近,与国外印度IND/Delhi/2016/05毒株进化关系最近。F基因核苷酸序列分析显示:试验毒株与国内北京China/BJ/2014毒株、吉林GZL-14毒株和浙江PPRV-FY毒株进化关系最近,与国外印度Izatnagar/94毒株和Izatnagar-94毒株进化关系最近。全基因及N、F基因核苷酸分析均显示试验毒株与西藏毒株及中国目前使用的疫苗毒株进化关系较远。N蛋白分析显示:试验毒株与中国河南CH/HNNY/2014毒株、CH/HNZM/2014毒株、中国广东GD/QY/2014毒株、中国浙江PPRV-FY毒株及国外的印度IND/Delhi/2016/05毒株进化关系最近。F蛋白氨基酸序列分析显示:试验毒株与河南CH/HNZK/2014毒株、北京China/BJ/2014毒株、吉林GZL-14毒株和浙江PPRV-FY毒株及国外印度Izatnagar/94毒株进化关系最近。N、F蛋白分析显示试验毒株与西藏毒株及中国目前使用的疫苗毒株进化关系较远。全基因及N、F基因核苷酸进化树构建结果均显示试验毒株与中国第二次疫情流行毒株同属第Ⅳ系的一个小分支,与国外的印度毒株进化关系较近。通过本研究表明,新疆阿克苏地区的本次疫情为PPRV感染绵羊所致,通过基因序列分析表明本试验毒株与中国2013~2014年PPR大流行时期的毒株进化关系最高,系统进化关系显示此次疫情毒株与中国2013~2014年PPR大流行时期的毒株同处于一个分支(第Ⅳ谱系)。本研究结果不仅能为该地区PPR诊断提供参考,而且能有效的为该地区的疫情防控提供理论依据。
[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.
【学位授予单位】：塔里木大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.26

【相似文献】