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1.
将2007年7月-2008年10月从吉林省猪群中分离到的3株H3N2亚型流感病毒,分别命名为A/swine/Jilin/A/2007(Sw/Jilin/A/07),A/swine/Jilin/B/2007(Sw/Jilin/B/07)和A/swine/Jilin/C/2008(Sw/Jmn/C/08).结合已知猪流感病毒(SIV)内部基因序列,设计合成6时特异性引物,通过RT-PCR技术分别扩增出6段内部基因,并与GenBank中相关序列进行比较,分析3株流感病毒内部基因的遗传进化关系.结果表明:在PB2、PB1、PA和NP的进化树上,3株H3N2分离株处于近代人源谱系内;在M和NS进化树中,Sw/Jilin/A/07和Sw/Jilin/B/07处于近代人源谱系内,而Sw/Jilin/C/08与禽源谱系内的H5亚型病毒亲缘关系最近.  相似文献   

2.
从天津地区不同猪场分离到6株H1N1亚型猪源流感病毒（SIV）。根据GenBank发表的H1N1亚型 SIV的核蛋白（NP）、基质蛋白（M）及非结构蛋白（NS）基因序列,分别设计3对引物,将RT-PCR产物克隆至pMD18-T载体,进行测序分析。遗传进化分析结果表明：A/swine/Tianjin/TJ2/2005（H1N1）与A/swine/Tianjin/TJ4/2006（H1N1）的NP、M及NS基因核苷酸序列在遗传进化树中均与A/swine/Guangdong/33/2006（H1N1）位于同一分支上,属于古典型H1N1猪谱系;A/swine/Tianjin/TJ3/2006（H1N1）与A/swine/Tianjin/TJ8/2006（H1N1)的NP、M及NS基因核苷酸在遗传进化树中均与A/Dunedin/2/2000（H1N1）组成一个大分支,可能起源于人谱系;A/swine/Tianjin/TJ6/2009（H1N1）与A/swine/Tianjin/TJ7/2009（H1N1）NP、M及NS基因核苷酸序列在遗传进化树中均与A/swine/Jiangsu/s15/2011（H1N1）位于同一分支上,属于类禽H1N1猪谱系。本试验对6株H1N1亚型SIV的NP、M及NS全基因序列进行分析,在一定程度上揭示了天津地区H1N1亚型SIV的基因进化与流行情况。  相似文献   

3.
为对2012年广州规模化养猪场疑似流感病料中分离鉴定的2株猪流感病毒A/swine/Guangdong/1519/2012(H3N2)(简称GD1519)和A/swine/Guangdong/1520/2012(H3N2)(简称GD1520)进行遗传进化及分子特征分析,对两株病毒进行了全基因组的测序分析。通过对其8个基因片段的基因来源进行分析,构建系统进化树,遗传进化分析结果显示GD1519 HA、PB2、PA和NP基因片段均来源于近期人源H3N2亚型Moscow/99-like亚分支;其NA和PB1基因片段来源于早期人源H3N2亚型Victoria/75-like亚分支;其M和NS基因片段来源于近期人源H3N2亚型New York/99-like亚分支。GD1520病毒的HA基因片段来源于近期人源H3N2亚型Moscow/99-like亚分支;其NA和PB1基因来源于近期人源H3N2亚型New York/99-like亚分支;其PB2、PA、NP、M和NS基因片段均来源于早期人源H3N2亚型Victoria/75-like亚分支。分析结果暗示2株病毒可能均由不同时期的类人源H3N2亚型流感病毒间基因重排而产生的新型H3N2亚型猪流感病毒。本研究结果进一步证明猪可能是不同来源流感病毒的基因"混合器",开展猪流感病毒相关分子流行病学研究具有重要的公共卫生意义。  相似文献   

4.
对3株H1N2亚型猪流感病毒(SIV):Sw/GX/17/05、Sw/HN/1/05和Sw/GX/13/06的血凝素(HA)、核蛋白(NP)、神经氨酸酶(NA)、基质蛋白(M)和非结构蛋白(NS)基因进行克隆和序列分析.结果显示:3株分离毒株HA、NP、NA、M和NS基因之间核苷酸同源性分别为91.3%～98.0%、98.4%～98.8%、97.4%～98.3%、98.8%～99.8%和98.1%～98.4%.遗传进化分析显示:分离毒株与美国分离的三源基因重排H1N2 SIV具有较近的亲缘关系;在HA、NP、M和NS基因进化树中,3株分离毒株均位于古典H1N1亚型SIV群,在NA基因进化树中,3株分离毒株则位于人流感病毒群.HA和NA基因推导氮基酸序列分别与代表毒株古典H1N1 SIV A/swine/Maryland/23239/1991(H1N1)和人H3N2流感病毒A/Buenos Aires/4459/96(H3N2)比较分析显示:HA(95.4%～96.1%)和NA(96.6%～97.2%)具有较高的氨基酸同源性;糖基化位点、抗原位点和受体结合位点(HA)处氨基酸存在一定的差异,这些氨基酸差异对病毒生物学特性的影响有待于进一步研究.  相似文献   

5.
本研究2012年底从辽宁省某屠宰场猪鼻咽拭子样品中分离到1株流感病毒,经HA—HI试验和RT—PCR鉴定为H1N1亚型猪流感病毒株,命名为A／swine/Liaoning／01／2012（H1N1）,通过对病毒的8个基因片段克隆并测序,并利用分子生物学软件进行遗传演化分析。结果表明,分离株HA基因裂解位点附近的氨基酸序列为IPSIQSRjG,符合低致病力流感病毒的分子特征。全基因组进化树结果表明,分离株的8个基因片段与A／swine／Jiangsu／40／2011（H1N1）株核苷酸同源性最高,分离株处在类禽型H1N1亚型遗传进化分支上;由于类禽型H1N1猪流感病毒具有潜在感染人的潜力,在国外和国内均有感染人的报道,因此,辽宁省首次分离到该型猪流感病毒对全省养猪业和公共卫生安全具有重要意义,值得深入研究。  相似文献   

6.
从山东地区疑似流感发病猪分离到 10株流感病毒 ,经国家流感中心鉴定均为 A型流感病毒 H9N2亚型。将其中 1株 Sw/ SD/ 1/ 2 0 0 3(H9N2 )的血凝素基因 (HA)、核蛋白基因 (NP)和非结构蛋白基因 (NS)进行克隆与测序 ,与Gen Bank收录的其他猪流感和禽流感 H9N2亚型的相关基因进行比较 ,推测 Sw/ SD/ 1/ 2 0 0 3(H9N2 )可能源于禽流感病毒 H9N2亚型和 H5 N1亚型的重组病毒 ;Sw/ SD/ 1/ 2 0 0 3的 HA氨基酸裂解位点与其他 H9N2亚型不同 ,Sw/ SD/1/ 2 0 0 3的 HA氨基酸裂解位点是 R- S- L- R- G,而其他猪流感和禽流感 H9N2亚型都是 R- S- S- R- G。  相似文献   

7.
为明确3株不同源性的H9N2亚型禽流感病毒(AIV)A/Chicken/Jilin/22/13(简称JL22)、A/Chicken/Jilin/24/13(简称JL24)、A/Duck/Jilin/37/13(简称JL37)基因组的遗传变异情况,本试验采用RT-PCR技术,分别扩增出3株AIV的8个基因片段,克隆后进行序列测定。结果显示,3株H9N2亚型AIV的主要致病基因均属于经典的欧亚种系。氨基酸比对发现JL22的HA氨基酸序列与A/Chicken/Hong Kong/G9/97和A/Duck/Hong Kong/Y280/97的HA氨基酸序列相比,在551位多了1个潜在糖基化位点。JL22、JL24、JL37的HA序列,在226位的氨基酸残基均为Leu,具有同哺乳动物唾液酸受体结合的特性,说明对人的感染性增强。M基因在31位上均发生了Asn取代Ser的现象,说明这些病毒对金刚烷胺产生了耐药性。由系统进化树可知3株毒株亲缘关系较远,各个基因所属分支也不具有统一性,且部分基因分别与鸡源、鸭源和猪源3种源性流感病毒株高度同源,推测这3株毒株是不同动物不同毒株经过长时间进化而发生自然重排的产物。  相似文献   

8.
从广东省不同猪场分离到4株H3N2亚型猪源流感病毒A/Swine/Guangdong/01/2004、A/Swine/Guang-dong/02/2004、A/Swine/Guangdong/03/2004、A/Swine/Guangdong/04/2004.根据GenBank公布的H3N2亚型猪源流感病毒的HA基因序列,设计1对引物,运用RT-PCR方法扩增四株病毒的HA基因,并进行测序和分析.同源性分析和遗传进化分析表明本实验的4株H3N2亚型SIV HA基因核苷酸序列同源性为99.8%～99.9%,在遗传进化树中均位于同一分支上.与参考毒株的比较分析表明,4个毒株与WHO推荐的2001-2004年北半球H3N2亚型流感疫苗株A/Moscow/10/99 HA基因的核苷酸序列同源性最高为99.4%～99.5%,4个毒株与A/Moscow/10/99 HA基因在遗传进化树中位于同一个小分支上.氨基酸序列比较发现,4个毒株HA基因裂解位点处的氨基酸序列均为PEKQTR↓G,4个毒株推导的氨基酸序列中均有11个糖基化位点,4个毒株HA蛋白226位受体结合位点(RBS)处氨基酸均为异亮氨酸(Ⅰ).4个毒株HA基因的氨基酸序列、受体结合位点以及糖基化位点均与A/Moscow/10/99相应的氨基酸序列一致.本试验的4株H3N2亚型猪源流感病毒的HA基因属于以A/Moscow/10/99为代表的近代类人H3N2亚型流感病毒,在一定程度上揭示了广东省H3N2亚型猪流感病毒HA基因进化与流行情况.  相似文献   

9.
为了解广东猪流感病毒(SIV)的流行变异情况,2010年11月从广东某规模猪场采集流感症状的猪鼻拭子60份,接种10日龄SPF鸡胚,分离到1株猪流感病毒,通过流感分型RT-PCR和HI试验鉴定为H3N2亚型SIV,命名为A/swine/Guangdong/L22/2010(H3N2),进行全基因序列测定及相似性分析发现,该分离株有低致病性流感分子特征,该毒株的8个基因片段连同最近广东猪群流行的流感毒株与2000年前后H3N2人流感病毒有较高的同源性.系统遗传演化显示,该病毒分离株可能是由1999年人源H3N2流感病毒A/Moscow/10/99(H3N2)进化而来.  相似文献   

10.
新型猪源性H1N1流感病毒能引起人和猪的呼吸道传染性疾病,自2009年4月起在全球范围内暴发,引起广泛的关注和研究,本试验拟对其分离株的气源性传播特点进行研究。2011年1月,华东某地区出现流感疫情,本研究从病死猪的鼻腔棉拭子和肺脏中分离到1株新型猪源性H1N1流感病毒A/swine/Shandong/07/2011;用荧光定量PCR方法检测发病猪场舍内、外的空气样品中病毒含量;建立气溶胶传染模型来分析该株病毒在实验条件下气源性传播的特点。结果显示:猪舍内空气样品的阳性率为26.10%,病毒含量在3.14～5.72log10copies.m-3空气之间;舍外下风向10m处空气样品的阳性率为40.70%,病毒含量在2.24～3.77log10copies.m-3空气之间;在传染模型中,A/swine/Shandong/07/2011能够造成气溶胶感染组试验猪的感染,但感染率比直接接触组低。研究表明,A/swine/Shandong/07/2011株具有形成病毒气溶胶的能力,在实验条件下能够引起气源性感染。  相似文献   

11.
H3亚型禽流感病毒分离株与变异株的生物学特性   总被引：5，自引：0，他引：5  
本研究对分离到的Ｈ３Ｎ８禽流感病毒株（分离株Ａ／Ｃｈｉｃｋ／Ｊｉｌｉｎ／９８,以下简称Ｊ１株）和传代过程中发生变异的变异株（Ａ／Ｃｈｉｃｋ／Ｊｉｌｉｎ／９９,以下简称Ｊ２株）进行了生物学和病理学特性的初步研究,发现在相同的时间内,Ｊ１株病毒致死鸡胚数量明显高于Ｊ２株病毒,病毒ＨＡ的滴度也明显高于Ｊ２;Ｊ１株的ＩＰＶＩ为１．４４,而Ｊ２为０．６４;毒株的致病性试验与ＳＰＦ感染鸡的器官变化表明Ｊ１组的病理学变化较Ｊ２组明显,且主要的嗜器官为肺、肝、法氏囊、胸腺、脾等主要的免疫器官;本研究还对ＳＰＦ感染鸡的病变器官进行了病原学检测、病理组织检测和电镜观察。结果表明两者存在较明显的差异。  相似文献   

12.
Swine influenza virus (SIV) of H1N1 and H3N2 subtypes are dominated in European pigs population. "Classical swine" H1N1 subtype was replaced by "avian-like" H1N1 subtype. It co-circulates with H3N2 reassortant possessing "avian" genes. In the present study, 41 SIV strains isolated from pigs with pneumonia, raised in 20 Polish farms, were identified and characterised. Since it was evidenced that isolates from the same geographic district and the same year of isolation are in 100% similar, 15 strains representing different district and different year of isolation were chosen to construct phylogenetic trees. Two genes, conservative matrix 1 (M1) and the most variable, haemagglutynin (HA), were sequenced and subjected into phylogenetic analysis. The results of the analysis confirmed that "avian-like" swine H1N1 strains evolved faster than classical SIV strains. HA gene of these isolates have been derived from contemporary strains of "avian-like" SIV. In contrast, the M1 gene segment may have originated from avian influenza viruses. H3N2 strain is located in swine cluster, in the main prevalent European group of H3N2 isolates called A/Port Chalmers/1/73-like Eurasian swine H3N2 lineage, which has evolved separately from the human H3N2 virus lineage around 1973.  相似文献   

13.
As well as H5 highly pathogenic avian influenza viruses (HPAIV), H7 HPAIV strains have caused serious damages in poultry industries worldwide. Cases of bird-to-human transmission of H7 HPAIV have also been reported [11]. On the outbreak of avian influenza, rapid diagnosis is critical not only for the control of HPAI but also for human health. In the present study, a rapid diagnosis kit based on immunochromatography for the detection of H7 hemagglutinin (HA) antigen of influenza A virus was developed using 2 monoclonal antibodies that recognize different epitopes on the H7 HAs. The kit detected each of the tested 15 H7 influenza virus strains and did not react with influenza A viruses of the other subtypes than H7 or other avian viral and bacterial pathogens. The kit detected H7 HA antigen in the swabs and tissue homogenates of the chickens experimentally infected with HPAIV strain A/chicken/Netherlands/2586/03 (H7N7). The results indicate that the present kit is specific and sensitive enough for the diagnosis of HPAI caused by H7 viruses, thus, recommended for the field application as a pen-site test kit.  相似文献   

14.
The H3N2 subtype of influenza A viruses isolated from pigs in the United States and Canada has shown both genetic and antigenic diversity. The objective of this study was to determine the serologic and genetic characteristics of contemporary strains of these viruses. Genetic analysis of 18 reference strains and 8 selected strains demonstrated differences in 1% to 9% of the nucleotides of the hemagglutinin (HA) gene. Phylogenetic analysis of the HA gene revealed 3 genetic clusters, as well as divergence of cluster III viruses from a cluster III prototype virus (A/Swine/Illinois/21587/99). By means of 1-way cross-hemagglutination inhibition with antiserum against 5 field isolates and 3 vaccine viruses, most of 97 isolates tested could be placed in 1 of 3 serogroups. The several isolates that did not react with any antiserum were in genetic cluster III, which suggests that continuous antigenic drift in cluster III may have resulted in virus variants. The efficacy of commercial vaccines against these virus variants should be evaluated with vaccination and challenge studies.  相似文献   

15.
为了解吉林省猪伪狂犬病毒（PRV）的遗传变异情况,本研究将采集自吉林省疑似发生猪伪狂犬病的临床样品,通过细胞传代、PCR检测和测序分析进行病毒分离鉴定。TCID50法测定分离毒株的病毒滴度,通过家兔感染试验测定分离毒株对家兔的致病性。对TK、gB、gC、gD和gE基因进行PCR扩增和测序,分析其遗传变异情况。结果表明,临床样品经PK15细胞传代后,有3份样品出现细胞病变,经PCR鉴定和测序分析表明,分离获得3株PRV,分别命名为PRV JL03、JL12和JL15株。通过PK15细胞测定分离毒株的病毒滴度分别为106.5、106.5和107.5TCID50/mL。6只家兔分别接种3株分离毒株（103.5TCID50/只）后均表现为体温升高、注射部位奇痒和四肢麻痹等症状,且于病毒接种后3~4 d全部死亡。3株分离病毒TK、gB、gC、gD和gE基因推导氨基酸序列分析结果表明,与参考毒株相比,分离株TK基因未发生氨基酸变异;gB、gC、gD和gE基因部分位点发生氨基酸缺失、插入或突变,其中gE基因在第48和496位分别存在1个天冬氨酸的插入,与国内报道的流行毒株变异特征一致。遗传进化树分析结果表明,3株分离毒株TK、gB、gC、gD和gE基因与国内2012年以后分离的参考毒株JS-2012和ZJ01等的上述基因均表现出较高的同源性,说明毒株间亲缘关系较近,位于同一分支;与国外分离毒株NIA3和Becker等亲缘关系较远,位于不同的分支;而与国内早期流行的毒株SC和Ea等亲缘关系介于二者之间。本研究成功分离鉴定了3株PRV变异株,分离毒株对家兔具有较强的致病性,该结果为吉林省PRV流行病学研究提供了新的数据,并为相关后续研究奠定基础。  相似文献   

16.
1,268 sera collected from slaughtered pigs in Hassia (FRG) from 1986 to 1988 were tested for antibodies against porcine and human influenza A virus strains using the single radial haemolysis test (SRHT). Antibodies against the porcine strains (subtype H1N1) A/Swine/Arnsberg/1/81, A/Swine/Iowa/15/30 and A/New Jersey/7/76 were detected in 411 (32.4%), 318 (25.1%) and 304 (24.0%) of sera, respectively. Up to 1988 a slight increase (10%) in the seroprevalence to A/Swine/Arnsberg/1/81 was noticed, whereas the results obtained with the other strains showed little variation. Antibodies against the human H1N1 strain A/Singapore/6/86 were only found in sera collected 1987 and 1988 in rates of 1.6% and 3.0%. Serological indication of infections with the human H3N2 strains A/Victoria/1/75, A/Hong Kong/1/68 and A/Philippines/2/82 could be shown in 286 (22.6%), 178 (14.4%) and 135 (10.6%) of the serum samples. Within the three year period the rate of sera positive for antibodies against A/Philippines/2/82 increased from 6.5% to 23.0%, whereas no variation in the rates were found using the other H3N2 strains. Antibodies simultaneously against porcine (H1N1) and human (H3N2) virus strains were detected in 9.9% of all sera tested.  相似文献   

17.
Genetic reassortment between human and avian influenza viruses can create pandemic viruses. Influenza surveillance of pigs in Jilin Province, in China during 2007–2008 revealed that there were two distinguishable genotypes: a human-like H3N2 genotype and a double-reassortant genotype derived from the human H3N2 and avian H5 viruses. In this study, viral infection potential, replication kinetics, and pathogenicity were compared. The solid-phase binding assay demonstrated that both viruses prominently maintained a preference for the human-type receptor and the reassortant A/swine/Jilin/37/2008 (Sw/JL/37/08) showed relatively higher binding affinities than the non-reassortant A/swine/Jilin/19/2007 (Sw/JL/19/07). Replication kinetics showed that Sw/JL/37/08 had higher replicability in MDCK cells than Sw/JL/19/07. The mouse experiments clearly revealed that Sw/JL/37/08 had higher virulence than Sw/JL/19/07 as measured by more significant body weight loss, higher viral lung load, delayed viral clearance from lungs, and more severe pulmonary lesions. Sequence analysis indicated that the absence of glycosylation sites at residue 126 of HA and 93 of NA, as well as the characteristic NS1 C-terminal PL residues of ESEV may account for the increased replication and pathogenicity of Sw/JL/37/08. These results may imply that human may have infection risk by the reassortant swine influenza virus and emphasize the necessity for enhanced viral surveillance strategies, which monitor reassortment events in nature to reduce the public health threat posed by influenza viruses with the potential for human-to-human transmission currently circulating in pig populations.  相似文献   

18.
Since the emergence of the 2009 pandemic (H1N1) virus (2009/H1N1) in April 2009, cases of transmission from humans to pigs have been reported frequently. In our previous studies, four 2009/H1N1 variants were isolated from pigs. To better understand the phenotypic differences of the pig isolates compared with the human isolate, in this study mice were inoculated intranasally with different 2009/H1N1 viruses, and monitored for morbidity, mortality, and viral replication, cytokine production and pathological changes in the lungs. The results show that all isolates show effective replication in lungs, but varying in their ability to cause morbidity. In particular, the strains of A/swine/Nanchang/3/2010 (H1N1) and A/swine/Nanchang/F9/2010 (H1N1) show the greatest virulence with a persisting replication in lungs and high lethality for mice, compared with the human isolate A/Liaoning /14/2009 (H1N1), which shows low virulence in mice. Furthermore, the lethal strains could induce more severe lung pathological changes and higher production of cytokines than that of other strains at an early stage. Amino acid sequence analysis illustrates prominent differences in viral surface glycoproteins and polymerase subunits between pig isolates and human strains that might correlate with their phenotypic differences. These studies demonstrate that the 2009/H1N1 pig isolates exhibit heterogeneous infectivity and pathogencity in mice, and some strains possess an enhanced pathogenicity compared with the human isolate.  相似文献   

19.
H3N2亚型猪流感病毒HA基因序列测定及抗原性分析   总被引：2，自引：3，他引：2  
采用RT-PCR技术对4株H3N2亚型猪流感病毒的HA基因进行了扩增,将获得的PCR产物分别与pMD18-T克隆载体连接,进行序列测定。测序结果显示,4个毒株均含有完整的开放阅读框,并且均未发现核苷酸插入或缺失现象;分离毒株间核苷酸同源性为99.4%~99.7%,氨基酸同源性为98.2%~99.3%。同源性分析表明,4个毒株与2003年的猪流感病毒广东分离株有很高同源性(均在99%以上),说明近段时间我国H3N2亚型的猪流感病毒变异不大,重组的频率不是很高,同时又与人流感病毒香港分离株有较高的同源性(均为99.4%)。交叉血凝抑制试验显示,S3株与其他3毒株抗原性差异明显。鉴于猪在流感病毒传播与复制间的特殊地位,应密切监测猪流感。  相似文献   

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