共查询到20条相似文献，搜索用时 406 毫秒
1.
旨在进一步了解山东省猪流感的流行情况及其病原特征,笔者于2019年春季,在山东省泰安某屠宰场采集130份猪鼻拭子,进行病毒分离鉴定;并对分离病毒进行全基因组测序和分子特征分析;用禽H9N2亚型标准抗原联合血凝抑制方法检测2018—2019年从山东省8个地区猪场采集的1 527份猪血清样品中的猪流感病毒抗体。结果显示：分离到1株H9N2亚型流感病毒,命名为A/swine/Shandong/TA009/2019（H9N2）。分离病毒与A/environment-air/Kunshan/NIOSH-BL20/2018（H9N2）和A/environment-air/Kunshan/NIOSH-BL25/2018（H9N2）遗传关系最近,其基因片段的核苷酸相似性均在99.5%以上。分离病毒的HA和NA基因属于Y280-like分支,PB2和M基因属于G1-like分支,PB1、PA、NP和NS基因属于SH/F98-like分支。分离病毒HA蛋白裂解位点处的氨基酸序列为“PSRSSR/GL”,符合低致病性禽流感病毒的分子生物学特性。HA蛋白的216位为L,具有结合人源唾液酸α 2,6-Gal的能力。血清学分析结果显示,9份血清中H9N2抗体呈阳性,其总阳性率为0.59%。综上：本研究分离到1株猪源H9N2亚型流感病毒,并在猪血清中检测到H9N2抗体,提示应加强对猪流感的流行情况及其病原特征的持续监测。 相似文献
2.
旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。 相似文献
3.
为了解华南地区猪群中猪流感病毒(SIV)的流行及其遗传变异情况,本研究从2016年~2017年广东、广西等地猪群236份猪肺脏病料组织和143份鼻拭子样品中分离鉴定得到3株SIV,全基因组测序和遗传演化分析结果显示,3个分离株均属于H1N1亚型欧亚类禽分支SIV,并且均与pdm09分支病毒株发生了重组。HA蛋白分子特征分析结果显示,A/Swine/Guangxi/NK/2016 HA蛋白第23位糖基化位点发生了缺失。3265份血清样品抗体监测结果显示,欧亚类禽H1N1、pdm09 H1N1和H3N2 SIV的血清抗体阳性率分别为27.53%、20.98%和34.85%。另外,0.64%的(21份)血清样品为H9N2亚型流感病毒抗体阳性,并且猪群中不同亚型和不同分支SIV之间混合感染的情况非常普遍。猪群中流感病毒血清抗体监测结果显示,EA H1N1、pdm09和H3N2亚型SIV HI抗体滴度最高均可达到1:1280,而H9N2亚型HI抗体滴度最高为1:160,表明H9N2 AIV虽然可以感染猪,但对猪还不适应。各月份的血清抗体阳性率分析显示,SIV的流行具有季节性,冬季(11月、12月和1月份)的流行最为严重。本研究可为华南地区猪群SI防控及疫苗株的筛选提供参考依据。 相似文献
4.
Isolation and genetic characterization of avian origin H9N2 influenza viruses from pigs in China 总被引：7，自引：1，他引：6
Yu H Hua RH Wei TC Zhou YJ Tian ZJ Li GX Liu TQ Tong GZ 《Veterinary microbiology》2008,131(1-2):82-92
As pigs are susceptible to infection with both avian and human influenza A viruses, they have been proposed to be an intermediate host for the adaptation of avian influenza viruses to humans. In April 2006, a disease caused by highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) occurred in several pig farms and subsequently overwhelmed almost half of China with more than 2,000,000 cases of pig infection. Here we report a case in which four swine H9N2 influenza viruses were isolated from pigs infected by highly pathogenic PRRSVs in Guangxi province in China. All the eight gene segments of the four swine H9N2 viruses are highly homologous to A/Pigeon/Nanchang/2-0461/00 (H9N2) or A/Wild Duck/Nanchang/2-0480/00 (H9N2). Phylogenetic analyses of eight genes show that the swine H9N2 influenza viruses are of avian origin and may be the descendants of A/Duck/Hong Kong/Y280/97-like viruses. Molecular analysis of the HA gene indicates that our H9N2 isolates might have high-affinity binding to the alpha2,6-NeuAcGal receptor found in human cells. In conclusion, our finding provides further evidence about the interspecies transmission of avian influenza viruses to pigs and emphasizes the importance of reinforcing swine influenza virus (SIV) surveillance, especially after the emergence of highly pathogenic PRRSVs in pigs in China. 相似文献
5.
猪作为流感病毒异源毒株间发生基因重组的"混合容器",其呼吸道上皮细胞上同时存在着能够感染人（SA α-2,6-Gal）和禽（SA α-2,3-Gal）两种流感病毒的受体,具备产生新型流感病毒的潜力。在我们的前期研究中,连续两年（2013年和2014年）从南宁地区某个规模化养猪场当中分离获得了2株新型甲型流感病毒重配的H3N2亚型猪流感病毒（swine influenza viruses,SIVs）。为了解SIVs在同一地方的遗传进化规律,我们在2018年至2019年间对该猪场进行了持续的监测,并于2019年再次成功分离获得了2株H3N2亚型的三源重组毒株,命名为A/swine/Guangxi/JG13/2019（简称JG13/2019）和A/swine/Guangxi/JG20/2019（简称JG20/2019）。遗传进化分析表明其基因重配形式与2013年分离株A/swine/Guangxi/JGB4/2013（简称JGB4/2013）和2014年分离株A/swine/Guangxi/JG1/2014（简称JG1/2014）相同,表面基因HA和NA来源于类人H3N2谱系,内部基因NP、M、PA、PB1和PB2来源于2009年甲型H1N1大流感谱系（pdm/09H1N1）,NS基因来源于古典型H1N1谱系。此外,新分离株JG13/2019和JG20/2019同早期分离株JGB4/2013和JG1/2014 HA和NA基因的核苷酸相似性分别为95.3%~97.4%和93.9%~97.0%,内部基因（NP、M、PA、PB1和PB2）的核苷酸相似性为96.2%~98.1%,NS基因的核苷酸相似性为97.1%~97.6%。通过分析比较这些年代不同毒株之间的关键氨基酸位点差异,结果发现JG20/2019和JG13/2019的HA蛋白仍旧保持了与人型受体结合的分子特征（190D、226I和228S）,却也出现了V223I或P227S的新变化,JG13/2019的PA蛋白（R356K）和PB2蛋白（I588T）也与之前的毒株有所不同。这些位点上的氨基酸改变是否影响到病毒的致病能力、复制能力以及跨种间传播能力,有待今后进一步研究。历经6年,携带有pdm/09 H1N1多种内部基因片段（PB2、PB1、PA、M、NP）和类人表面基因（HA和NA）的H3N2亚型SIVs依旧在同一个猪场的猪群中流行,虽然其关键的功能区域出现了基因突变,但是仍然保持着能够感染人的受体结合特性。因此,加强对SIVs流行情况的监测,将为今后防控人类流感大暴发提供预警。 相似文献
6.
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
7.
采用RT-PCR技术对分离的H1N1亚型猪流感病毒的HA基因进行了扩增,将获得的PCR产物与pMD18-T载体连接,进行序列测定。同源性分析结果表明,分离毒株与其他H1N1亚型猪流感病毒的HA基因核苷酸同源性为70.7%～90.8%,与A/swine/Zhejiang/1/2007的同源性最高,与其他毒株的同源性相对较低。系统进化树分析结果表明,山东分离株的HA基因与欧洲谱系猪流感病毒进化关系最近,证明该分离株可能来源于北美谱系和欧亚谱系猪流感病毒的重组。 相似文献
8.
9.
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
10.
Genetic diversity of H9N2 influenza viruses from pigs in China: a potential threat to human health? 总被引：3，自引：0，他引：3
Yu H Zhou YJ Li GX Ma JH Yan LP Wang B Yang FR Huang M Tong GZ 《Veterinary microbiology》2011,149(1-2):254-261
Pandemic strains of influenza A virus might arise by genetic reassortment between viruses from different hosts. Pigs are susceptible to both human and avian influenza viruses and have been proposed to be intermediate hosts or mixing vessels, for the generation of pandemic influenza viruses through reassortment or adaptation to the mammalian host. In this study, we summarize and report for the first time the coexistence of 10 (A-J) genotypes in pigs in China by analyzing the eight genes of 28 swine H9N2 viruses isolated in China from 1998 to 2007. Swine H9N2 viruses in genotype A and B were completely derived from Y280-like and Shanghai/F/98-like viruses, respectively, which indicated avian-to-pig interspecies transmission of H9N2 viruses did exist in China. The other eight genotype (C-J) viruses might be double-reassortant viruses, in which six genotype (E-J) viruses possessed 1-4 H5-like gene segments indicating they were reassortants of H9 and H5 viruses. In conclusion, genetic diversity of H9N2 influenza viruses from pigs in China provides further evidence that avian to pig interspecies transmission of H9N2 viruses did occur and might result in the generation of new reassortant viruses by genetic reassortment with swine H1N1, H1N2 and H3N2 influenza viruses, therefore, these swine H9N2 influenza viruses might be a potential threat to human health and continuing to carry out swine influenza virus surveillance in China is of great significance. 相似文献
11.
在2019年1月-2019年6月对云南出现呼吸道疫病的57个鸡场进行H9亚型禽流感检测的基础上,选取石林和楚雄2个H9亚型禽流感阳性样品进行病毒分离。从分离的H9N2亚型禽流感病毒感染鸡胚尿囊液中提取总RNA,采用特异性引物经反转录PCR分别扩增HA和NA基因,PCR产物纯化后进行测序。序列比对及系统发育分析结果表明,云南2株H9N2毒株HA基因核苷酸序列同源性为94.2%,NA基因核苷酸序列同源性为93.6%,系统进化分析表明云南H9N2亚型禽流感病毒HA和NA基因均属于欧亚谱系中的类ADKHKY28097分支（Y280-like）,ACKYN12019和ACKYN72019 HA基因之间的同源性为94.3%,与参考毒株ACKJX2448的同源性最高,为95.6%~98.5%,与中国流行的H9N2代表株和疫苗株同源性较低。HA蛋白333-340位裂解位点为PSRSSR↓GLF,具有低致病性禽流感病毒分子特征,受体结合位点均发生E198T和Q234L的突变,具有人样受体结合特征,在29、141、298、305、313、492位氨基酸有6个糖基化位点。ACKYN12019和ACKYN72019 NA基因同源性为93.6%,与Y280-like代表毒株的同源性分别为97.1%~97.5%和93.7%~94.6%,NA蛋白缺失63、64、65位氨基酸,在44、69、86、146、200、234位氨基酸处存在6个潜在的糖基化位点,NA蛋白红细胞结合（HB）位点分析发现,368-369、399-403、432位氨基酸处存在变异。研究结果显示,H9N2亚型禽流感病毒一直处于不断的变异之中,故应加强其监测与防控。 相似文献
12.
为了解猪流感病毒(SIV)的变异情况,我们2009年11月从河北某养殖场采集呈流感症状的猪鼻拭子40份,接种10日龄SPF鸡胚,分离到一株猪流感病毒,通过RT-PCR和血凝抑制试验鉴定为H1N1亚型,命名为A/swine/Hebei/15/2009(H1N1),其全基因序列测定及同源性分析发现,8个基因片段均与2000年左右H1N1人流感病毒有较高的同源性。系统遗传演化显示,该病毒分离株是由2000年人源H1N1流感病毒A/Dunedin/2/2000(H1N1)进化而来。抗原性分析显示该株与甲型H1N1流感病毒和经典H1N1病毒株抗原性差异较大。对小鼠致病性试验表明该病毒株可以直接感染小鼠并导致小鼠轻微临床症状和组织病理学变化,但不致死小鼠,表现为低致病性。 相似文献
13.
14.
本研究从广东省某猪场采集37份疑似猪流感症状的猪鼻拭子样品,接种于9日龄SPF鸡胚并收集尿囊液,通过血凝试验、血凝抑制试验和RT-PCR鉴定,分离得到一株猪流感病毒,经RT-PCR分别扩增8个基因片段,进行基因测序及序列分析,与GenBank收录的参考毒株比对并构建进化树。结果显示,分离毒株为H1N1亚型猪流感病毒,将其命名为A/swine/Guangdong/2/2018（H1N1）。遗传进化分析显示,分离株8个片段的核酸序列与A/swine/Guangdong/L3/2009（H1N1）对应序列的同源性均达99%以上,与经典型H1N1亚型猪流感病毒处于同一分支。分离毒株HA的裂解位点为PSIQSR↓GL,符合低致病性流感病毒分子特征。HA基因受体位点为190D、225G和226Q,表明本毒株既可以结合SAα-2,6-Gal型人类流感病毒SA受体,也有结合SAα-2,3-Gal型禽类流感病毒SA受体的可能,在28、40、104、304、498、557位氨基酸处有6个潜在糖基化位点;NA蛋白在50、58、63、68、98、146、235位氨基酸处有6个潜在糖基化位点,NA蛋白氨基酸序列活性中心位点为119E、199D、223I、275H、293R、295N,氨基酸分析位点未出现突变,表明本分离株对神经氨酸酶抑制剂类药物的敏感性较高,但在M2蛋白中,31位氨基酸由敏感型的（S）突变为抗药的（N）,提示可能对金刚烷胺类药物产生耐药性。开展猪流感病毒分离鉴定与遗传进化分析将为广东地区的猪流感流行和变异情况提供重要信息。 相似文献
15.
Maldonado J Van Reeth K Riera P Sitjà M Saubi N Espuña E Artigas C 《Veterinary journal (London, England : 1997)》2006,172(2):377-381
This paper reports on a serological and virological survey for swine influenza virus (SIV) in densely populated pig areas in Spain. The survey was undertaken to examine whether the H1N2 SIV subtype circulates in pigs in these areas, as in other European regions. Six hundred sow sera from 100 unvaccinated breeding herds across Northern and Eastern Spain were examined using haemagglutination inhibition (HI) tests against H1N1, H3N2 and H1N2 SIV subtypes. Additionally, 225 lung samples from pigs with respiratory problems were examined for the presence of SIV by virus isolation in embryonated chicken eggs and by a commercial membrane immunoassay. The virus isolates were further identified by HI and RT-PCR followed by partial cDNA sequencing. The HI test on sera revealed the presence of antibodies against at least one of the SIV subtypes in 83% of the herds and in 76.3% of the animals studied. Of the 600 sow sera tested, 109 (18.2%), 60 (10%) and 41 (6.8%) had SIV antibodies to subtype H1N2 alone, H3N2 alone and H1N1 alone, respectively. Twelve H3N2 viruses, 9 H1N1 viruses and 1 H1N2 virus were isolated from the lungs of pigs with respiratory problems. The analysis of a 436 nucleotide sequence of the neuraminidase gene from the H1N2 strain isolated further confirmed its identity. Demonstrably, swine influenza is still endemic in the studied swine population and a new subtype, the H1N2, may be becoming established and involved in clinical outbreaks of the disease in Spain. 相似文献
16.
在实验室前期利用猪肾细胞（PK-15）CRISPR/Cas9全基因组敲除文库（PK-15-GeCKO）筛选到猪流感病毒（swine influenza virus,SIV）复制相关的宿主因子磷酸十二烷基磷酸酯-葡萄糖基转移酶（ALG5）的前提下,深入研究ALG5与SIV复制的关系。本研究利用CRISPR/Cas9技术构建了包含ALG5基因向导RNA（gRNA）的质粒LentiGuide-puro-ALG5,并通过慢病毒包装感染稳定表达Cas9蛋白的新生猪气管上皮（NPTr）细胞系,采用有限稀释法筛选ALG5基因敲除的单克隆细胞株,通过靶基因组测序及Western blot验证了ALG5基因在NPTr细胞上的敲除水平,通过CCK-8试验验证基因敲除细胞和野生型细胞的细胞活性。结合TCID50和Western blot试验检测了ALG5敲除和过表达对SIV复制的影响。同时,检测了SIV感染NPTr细胞后内源性ALG5蛋白表达量的变化。最后检测了ALG5敲除对猪流感毒株F26复制的影响。结果显示：获得了ALG5敲除的NPTr单克隆细胞系（ΔALG5）,ALG5基因敲除细胞与野生型细胞的细胞活性无显著差异。ALG5基因敲除显著抑制SIV的增殖,过表达促进SIV的增殖。在病毒感染条件下,随着病毒的增殖,ALG5的蛋白表达量上调。ALG5基因敲除后,也可以抑制猪流感毒株F26的复制,无毒株特异性。本研究表明,ALG5是影响猪流感病毒复制过程的一个重要宿主因子,为研究猪流感病毒的复制和致病机制奠定了基础。 相似文献
17.
X.‐H. Song H. Xiao Y. Huang G. Fu B. Jiang Y. Kitamura W. Liu D. Liu G. F. Gao 《Zoonoses and public health》2010,57(4):291-298
Several highly pathogenic H5N1 avian influenza viruses were isolated from swine populations in Fujian Province, China, since 2001. Because it is thought that H5N1 infection in pigs might result in virus adaptation to humans, we surveyed swine populations in Fujian Province in 2004 and 2007 for serological evidence of the infection. Twenty‐five pig farms covering all nine administrative districts of Fujian Province were sampled and a total of 1407 serum specimens were collected. The haemagglutination inhibition (HI) tests revealed no evidence of H5 infection and only a few cases of H9 infection. The negative results for H5 infection were further verified by micro‐neutralization tests. By contrast, H1 influenza virus infections were prevalent in swine in both surveys according to the results of enzyme‐linked immunosorbent assay (ELISA). The H3 infection rate was reduced dramatically in 2007 compared with 2004, when examined by HI and ELISA. In summary, the results imply that the swine populations in Fujian Province had not been affected greatly by the H5N1 avian influenza virus, given that there is no serological evidence that H5N1 influenza virus has infected the pig populations. The reported isolates represent only sporadic cases. 相似文献
18.
本试验针对猪流感病毒（swine influenza virus,SIV）NP基因保守区域设计并合成6条引物,建立了SIV的环介导等温扩增（LAMP）检测方法,并进行了特异性、敏感性和重复性试验。结果显示,该方法可特异性检测H1N1、H1N2、H3N2、类禽H1N1亚型SIV及甲型H1N1流感病毒,但不能检测猪繁殖与呼吸综合征病毒、猪细小病毒、猪瘟病毒、猪伪狂犬病病毒、猪圆环病毒2型、日本乙型脑炎病毒;该方法的最低检测量为100拷贝/μL质粒DNA。结果表明建立的LAMP方法具有较高的敏感性和特异性,可用于SIV的快速检测。 相似文献
19.
试验旨在确定H9N2亚型猪源性流感病毒（SIV）在小鼠肺微血管内皮细胞（PMVEC）中增殖的最佳条件。将PMVEC解冻、复苏、培养,取长成单层的PMVEC,在不同浓度TPCK-胰蛋白酶维持液（0.1、0.2、0.3、0.4、0.6、0.8、1.0 μg/mL）、不同H9N2亚型SIV（A/swine/HeBei/012/2008/（H9N2）接种剂量（1：10、1：100、1：1 000、1：10 000、1：100 000、1：1 000 000和1：10 000 000）及不同病毒吸附时间（0.5、1.0、2.0和3.0 h）条件下观察PMVEC形态变化,并测定细胞上清液中H9N2亚型SIV的HA滴度。在未加病毒的情况下,低于0.6 μg/mL的TPCK-胰蛋白酶对PMVEC的生长未造成任何影响,但随着TPCK-胰蛋白酶浓度的增加,PMVEC开始出现肿胀、变圆,甚至脱落;采用含有0.6 μg/mL TPCK-胰蛋白酶维持液将H9N2亚型SIV稀释为不同的浓度感染PMVEC,在0.3 μg/mL TPCK-胰蛋白酶维持液、10-2病毒稀释倍数感染条件下48和72 h HA滴度分别为4.6log2和6.4log2;病毒吸附时间为2 h且中间震荡20 s的条件下H9N2 HA滴度最佳。结果表明,当TPCK-胰蛋白酶维持液浓度为0.3 μg/mL、病毒接种浓度为10-2、吸附时间为2 h且中间震荡20 s时,H9N2亚型SIV在PMVEC中增殖最佳,达到6.8log2。 相似文献
20.
猪流感病毒H1、H3、N1、N2亚型分型 RT-PCR方法的建立 总被引：1，自引：0，他引：1
根据GenBank中H1N1和H3N2亚型猪流感病毒(SIV)血凝素(hemagglutinin,HA)、神经氨酸酶(neuraminidase,NA)和M基因保守序列,分别设计合成了5对特异性引物,利用RT-PCR技术对SIV的型和亚型进行鉴定。结果表明,该方法的型RT-PCR可以检测出104 EID50病毒量所提取的RNA;H1、H3、N1和N2的亚型RT-PCR均可以检测出104 EID50病毒量所提取的RNA。除每对特异性引物所对应的亚型外,对其他亚型及猪繁殖与呼吸综合征病毒(PRRSV)和猪瘟病毒(CSFV)的检测均为阴性,应用该方法对临床样品进行检测,其结果与病毒分离结果符合率为100%。结果表明,该方法特异性好、敏感性高,有望成为SIV的一种特异、敏感、快速的分型检测方法,为猪流感分子流行病学的调查奠定了良好的基础。 相似文献