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1.
Jaime Maldonado Pere Riera Marta Sitja Narcis Saubi Enric Espuna Carlos Artigas 罗天海 《国外畜牧学(猪与禽)》2006,26(6):35-38
本文报道了对西班牙养猪密集地区进行的猪流感病毒(SIV)血清学和病毒学调查结果,其目的是检测H1N2亚型猪流感病毒是否和在欧洲其它地区一样也流行于这些地区。在西班牙北部和东部地区100个未接种猪流感疫苗的繁育群采集了600份母猪血清,以进行抗H1N1、H3N2和H1N2亚型病毒的血凝抑制试验(HI);再借助鸡胚病毒分离法和商品薄膜免疫分析法,对采自有呼吸道疾患的225份病猪肺样本进行检测以确定是否存在猪流感病毒。通过HI和逆转录酶-聚合酶链式反应(RT-PCR)方法,再辅以cDNA部分序列的测定,对分离毒株进行鉴定。对血清所作的HI试验表明,在83%受测猪群和76·3%受测猪中至少存在抗一种猪流感病毒亚型的抗体。在受检的600份母猪血清中,仅含抗H1N2、H3N2或H1N1亚型病毒抗体的样本数分别为109份(占总样本比例的18·2%)、60份(10%)和41(6·8%)。从有呼吸道疾患的猪肺样本中分离到12株H3N2亚型病毒、9株H1N1亚型病毒和1株H1N2病毒。对H1N2亚型分离株的神经氨酸酶基因436个核苷酸序列进行的分析,进一步证实了其身份。显然,猪流感依然流行于所研究猪群中,同时一种新的亚型病毒(即H1N2亚型病毒)可能正在开始流行,并且也在西班牙引起了临床发病。 相似文献
2.
为了解广西猪群中猪流感(SI)的流行情况,以猪流感病毒H3N2亚型作为诊断抗原,采用微量血凝抑制试验(HI)方法,对广西部分市(县)的猪群进行猪流感的血清学调查.在12个市(县)采集的1000份血清中,有10个市(县)检出H3N2抗体,共有257份血清呈H3N2抗体阳性,平均阳性率为25.7%.百色的阳性率最高,达到82.9%,贵港和南宁市郊两地没有检出阳性,其它9个市(县)的阳性率在13.0%～45.7%之间.结果表明我区猪群中很可能存在H3N2亚型猪流感感染,其中百色地区猪流感的感染率比较高. 相似文献
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为了对供港猪群中的猪流感流行情况进行分析,从华南地区供港猪群中用无菌棉拭子采集鼻腔粘液样品,采用鸡胚接种方法,从供港猪群中分离出了2株不同亚型的猪流感病毒株,经国家流感中心鉴定分别为H1N1和H3N2亚型。本研究设计了猪流感常见亚型的HA和NA分型特异性引物,建立了猪流感型特异性RT-PCR检测方法;对分离鉴定的2株猪流感病毒和禽流感H5N1 HI检测抗原进行了RT-PCR检测,并对其部分HA和NA基因进行克隆测序分析。对供港猪群的血清检测结果表明:供港猪群中H1N1和H3N2亚型抗体阳性率分别为26.87%、38.26%,禽流感H5N1和H9N2亚型抗体阳性率均为0%。 相似文献
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以H3亚型禽流感病毒(AIV)免疫BALB/c小鼠,取其脾细胞与SP2/0骨髓瘤细胞融合,用血凝抑制试验(HI)筛选阳性杂交瘤细胞并克隆化。结果获得了4株针对H3亚型禽流感病毒血凝素的单克隆抗体,分别命名为1D7、1F9、5A4、5F5。这些单克隆抗体小鼠腹水HI效价为212～214。用H1、H6、H10亚型禽流感病毒各2株,H4、H5、H9和新城疫病毒(NDV)各1株进行特异性试验。结果表明:所有这些单抗仅与H3亚型AIV发生特异性HI反应,而不与其他亚型AIV及NDV反应。用28株H3亚型禽流感病毒进行排谱试验,结果证明:4株单抗均具有广谱性,其中1F9、5A4、5F5与受试的28株H3亚型AIV均反应,而1D7只与其中的26株反应。以上单抗将为控制畜禽及人类的流感提供必需的诊断试剂。 相似文献
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《动物医学进展》2020,(6)
从北京市延庆区某疑似发生犬流感的德国牧羊犬采集犬鼻拭子,采用鸡胚接种、传代、HI试验、HA和NA基因扩增和序列同源性分析及比格犬回归试验等方法对病毒进行分离鉴定。结果表明,分离的病毒具有HA活性,且HA活性可被H3亚型禽流感病毒阳性血清所抑制(HI抗体效价为1∶160),与H1、H5、H7亚型禽流感病毒阳性血清和犬流感阴性血清HI试验均为阴性反应(HI抗体效价1∶10)。分离病毒HA基因、NA基因与A/canine/Georgia/89750.1/2017(H3N2)毒株HA、NA基因的同源性均为99.8%。攻毒后第4天开始,2只犬均出现体温升高、咳嗽、流鼻和精神沉郁等临床症状,并从鼻拭子中分离到病毒,该病毒是一株H3N2亚型的犬流感病毒,将其命名为A/canine/China/Huabei-170607/2017(H3N2)。 相似文献
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猪流感抗体间接ELISA检测方法的建立 总被引：12，自引：1，他引：12
猪流感病毒A／Swine／Fujian／668／2001（H3N2）株感染的鸡胚尿囊液，经差速离心后，再经蔗糖密度梯度离心，提纯、纯化的猪流感病毒经NP-40处理并反复冻融，作为猪流感间接ELISA抗原，确立了间接ELISA检测方法。对29份HI试验猪流感为阴性的血清进行了检测，经统计学分析，确定间接ELISA判定标准，被检血清OD490nm值≥0．20判定为阳性。该方法对猪瘟等11种猪疫病阳性血清无交叉反应，批内和批间重复试验的吸收变异系数分别在3．34％～8．12％和6．2％～9．04％之间。与HI的符合率达到92．8％，经卡方检验（P〈0．01）比HI试验敏感。为猪流感抗体检测提供了快速、准确、简便的方法。 相似文献
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到目前为止,意大利已经分离到了H1N1、H1N2和H3N2 3个亚型猪流感病毒。2006年,在一例有咳嗽症状的猪上分离到了一种新的猪流感病毒亚型(H3N1)。通过对肺组织的RT-PCR检测,对试验猪人工感染该新亚型流感病毒及利用空斑试验对该毒株的克隆鉴定,证实了这一独特的H和N组合的猪流感病毒的存在。同时,对该新型毒株进行了抗原及遗传特性鉴定。基因系统发育分析结果表明,H3N1亚型的完整HA基因序列与3株意大利H3N2亚型具有高度的一致性。这3个意大利亚型中有一株分离于2001年,其它2株分离于2004年,而该毒株的NA序列全长与2004年于意大利分离得到的3株H1N1亚型猪流感病毒极其相似。其余的基因片段也分别与当前意大利流行的H1N1与H3N2猪流感病毒极为相似。这表明,该新亚型猪流感病毒可能是H1N1与H3N2亚型的一个重组体。 相似文献
10.
为了制备具有HI活性的HI亚型流感病毒特异性单克隆抗体(MAb),本研究以H1N1亚型猪流感病毒(SIV)株A/Swine/Guangdong/718/01(H1N1)为免疫原,免疫BALB/c小鼠,经常规细胞融合后,血凝抑制(HI)方法进行检测,融合细胞经稀释克隆纯化后,获得11株能稳定分泌抗血凝素特异性HI MAb的杂交瘤细胞株。鉴定表明,所获MAb与其他具有血凝活性的病毒以及其他14个HA亚型的流感病毒均不具有HI交叉反应,表明这11株MAb均具有良好的流感病毒亚型特异性。其中A6F、2BBF和2BB与其他H1亚型流感病毒分离株的HI试验证实我国不同地区分离株之间的抗原性存在一定差异。11株MAb对H1SIV抗原的HI试验结果显示其HI效价有明显差异。叠加实验表明这些MAb分别识别HA抗原的不同表位。间接免疫荧光试验表明,2BBF、8HB、1DH、7FC和2BB均可与2009年流行H1N1病毒A/California/04/2009HA抗原发生特异性反应。这些MAb特异性的研制为H1亚型流感病毒的疫情病原学快速诊断以及病毒抗原性变异的相关研究提供了物质基础。 相似文献
11.
Dana Skibbe En-Min Zhou Bruce H Janke 《Journal of veterinary diagnostic investigation》2004,16(1):86-89
A commercial indirect swine influenza virus (SIV) H1N1 enzyme-linked immunosorbent assay (ELISA) was compared with the hemagglutination inhibition (HI) assay by testing 72 samples from experimentally infected pigs and 780 field samples of undefined SIV status. The HI assay was performed using SIV isolates A/Swine/IA/73 for H1N1 and A/Swine/IA/8548-1/98 for H3N2. The ELISA used an SIV isolated in 1988. The results showed that HI and ELISA detected an antibody in 11 and 6, respectively, of 72 serum samples collected from pigs experimentally infected with a 1992 SIV isolate (A/Swine/IA/40776/92). The presence of antibodies in these experimental samples was confirmed by HI tests in which all 72 samples were positive against the homologous virus, a more recent H1N1 SIV isolate (A/Swine/NVSL/01) supplied by National Veterinary Services Laboratories, Ames, Iowa, and a 1999 H1N1 isolate currently used in a commercial vaccine. On testing 780 field samples, an overall agreement of 85.5% was generated between the HI and ELISA. This study demonstrated that the ELISA is a useful serodiagnostic screening test at herd level for detecting swine antibodies against SIV. However, a new SIV isolate representing current SIV strains circulating in the field is needed to replace the older isolates used in the HI and ELISA to increase the test accuracy for serodiagnosis of SIV. 相似文献
12.
N Masurel G.F de Boer W.J.J Anker A.D.N.H.J Huffels 《Comparative immunology, microbiology and infectious diseases》1983,6(2):141-149
In the period December 1979-May 1980 a respiratory disease spread rapidly through pig herds in The Netherlands. Surveillance of 12 pig farms resulted in isolation of 22 influenza A-Swine-H1N1 (Hsw1N1) strains from 9 pig herds. The morbidity rate was high but the mortality rate was nil. Retardation in growth was observed. Sera collected from affected pig herds showed a fourfold increase in haemagglutination inhibition (HI) titre against A-Swine-H1N1 virus. Sera collected on five farms showed a geometric mean HI titre against the A-H3N2 virus above 100. A significant HI titre increase against this virus was found in sera collected on three farms. These findings indicated a recent infection by this virus. A-H3N2 virus was not isolated. The Dutch Swine-1980 isolates showed in the cross-HI test a distant antigenic relationship with the classical A/Swine/Iowa/30 (H1N1) virus and one-sided close antigenic relationship with A/New Jersey/76 (H1N1) virus.HI antibody to A/Swine/Nederland/80 (H1N1) virus was found in 4, 0, and 44%, to A/New Jersey/76 (H1N1) virus in 0.5, 0.4, and 42%, and to A/Swine/Iowa/30 (H1N1) virus in 0.5, 1, and 30% of pig sera collected in 1976, 1977, and 1980, respectively. HI antibody to A/Hong Kong/68 (H3N2) virus was detected in 36, 56, and 68%, and to A/Victoria/75 (H3N2) virus in 38, 73, and 68% of these sera, respectively.The results of this study indicate that pigs in The Netherlands, like those in North America, Southeast Asia, Japan, and Western Europe harbour A-Swine-H1N1 and A-H3N2 influenza viruses and are thus potential reservoirs for future human pandemics. 相似文献
13.
禽流感病毒CH02株(H9)的鉴定及其NS 基因分子特征分析 总被引：2，自引：2，他引：0
从疑似感染H9亚型禽流感病毒的病鸡内脏组织中分离到1株能凝集鸡红细胞的病毒,通过血凝抑制试验、鸡胚中和试验、RT-PCR鉴定,确认其为H9亚型禽流感病毒,并命名为CH02株。该病毒HA效价为27.67±0.58;其血凝性可被抗H9亚型禽流感病毒阳性血清完全抑制,HI效价为7log2;鸡胚半数感染量(EID50)为10-8.68 EID50/0.1 mL;最小致死剂量致鸡胚死亡的平均时间(MDT)为85.6 h;1日龄鸡脑内致病指数(ICPI)为0.625;其NS基因与香港株、南京株、北京株、汕头株、韩国株、巴基斯坦株的核苷酸同源性为88.6%~100%,经进化分析CH02株与香港株(A/duck/Hong Kong/Y280/97)同属一个分支。 相似文献
14.
本研究由疑似猪流感病料样品中分离一株病毒,通过HA、HI、电镜观察、生物学特性测定及全基因序列测定表明,分离病毒株为H3N2亚型人流感病毒和H1N1亚型古典猪流感病毒(SIV)的重组体,命名为A/Swine/Fujian/F2/07(H3N2).该分离株含有8个基因片段,共13 442 bp,与GenBank中登录的H3N2亚型人流感病毒、H1N1亚型古典SIV和H3N2亚型SIV进行比较分析显示:分离株的HA、NS、NA基因与H3N2亚型人流感病毒株的同源性分别为84.7 ％～98.1％、94.4 ％～99.5％和88.6 ％～97.6％;与H3N2亚型SIV的核苷酸同源性分别为87.7％～98.5％、82.5 ％～99.9％和87.6 ％～98.4％;M基因与H3N2亚型人流感病毒和SIV的同源性均在90.1％以下,而与H1N1亚型古典SIV的同源性在97.6％以上.基因型分析表明分离株的PB2、PBI、PA、HA、NP、NA和NS基因片段来源于1975年～1982年的人流感病毒,而M基因来源于H1N1亚型古典SIV,充分证明猪作为流感病毒“混合器”的作用. 相似文献
15.
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
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X M Zhang W Herbst H Lange-Herbst T Schliesser 《Zentralblatt für Veterin?rmedizin. Reihe B. Journal of veterinary medicine. Series B》1989,36(10):765-770
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.
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
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Tiina Nokireki Taina Laine Laura London Niina Ikonen Anita Huovilainen 《Acta veterinaria Scandinavica》2013,55(1):69
Background
Swine influenza is an infectious acute respiratory disease of pigs caused by influenza A virus. We investigated the time of entry of swine influenza into the Finnish pig population. We also describe the molecular detection of two types of influenza A (H1N1) viruses in porcine samples submitted in 2009 and 2010.This retrospective study was based on three categories of samples: blood samples collected for disease monitoring from pigs at major slaughterhouses from 2007 to 2009; blood samples from pigs in farms with a special health status taken in 2008 and 2009; and diagnostic blood samples from pigs in farms with clinical signs of respiratory disease in 2008 and 2009. The blood samples were tested for influenza A antibodies with an antibody ELISA. Positive samples were further analyzed for H1N1, H3N2, and H1N2 antibodies with a hemagglutination inhibition test. Diagnostic samples for virus detection were subjected to influenza A M-gene-specific real-time RT-PCR and to pandemic influenza A H1N1-specific real-time RT-PCR. Positive samples were further analyzed with RT-PCRs designed for this purpose, and the PCR products were sequenced and sequences analyzed phylogenetically.Results
In the blood samples from pigs in special health class farms producing replacement animals and in diagnostic blood samples, the first serologically positive samples originated from the period July–August 2008. In samples collected for disease monitoring, < 0.1%, 0% and 16% were positive for antibodies against influenza A H1N1 in the HI test in 2007, 2008, and 2009, respectively. Swine influenza A virus of avian-like H1N1 was first detected in diagnostic samples in February 2009. In 2009 and 2010, the avian-like H1N1 virus was detected on 12 and two farms, respectively. The pandemic H1N1 virus (A(H1N1)pdm09) was detected on one pig farm in 2009 and on two farms in 2010.Conclusions
Based on our study, swine influenza of avian-like H1N1 virus was introduced into the Finnish pig population in 2008 and A(H1N1)pdm09 virus in 2009. The source of avian-like H1N1 infection could not be determined. Cases of pandemic H1N1 in pigs coincided with the period when the A(H1N1)pdm09 virus was spread in humans in Finland. 相似文献19.
Antigenic characterization of an H3N2 swine influenza virus isolated from pigs with proliferative and necrotizing pneumonia in Quebec. 总被引：2，自引：0，他引：2 
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下载免费PDF全文 M H Bikour E Cornaglia J M Weber Y Elazhary 《Canadian journal of veterinary research》1994,58(4):287-290
A new strain of swine influenza A virus, designated A/Swine/Saint-Hyacinthe/150/90 has been isolated from pigs with severe proliferative and necrotizing pneumonia in Quebec. The antigenic characterization of the hemagglutinin was performed by hemagglutination inhibition test, immunoblot and indirect immunoprecipitation using polyclonal antisera. Only the last test was able to detect an antigenic relationship between the hemagglutinin of this isolate and an H3 subtype influenza virus. The immunoprecipitation test was a useful alternative for determining the hemagglutinin of influenza A virus subtypes. The neuraminidase inhibition test demonstrated a reactivity between the A/Swine/Saint-Hyacinthe/150/90 and antiserum against a N2 subtype influenza virus. Our results indicate that this new strain isolated for the first time in the porcine population of Canada is related to A/Sw/Hong Kong/76 H3N2 swine influenza virus. 相似文献
20.
Influenza A is a respiratory disease common in the swine industry. Three subtypes, H1N1, H1N2 and H3N2 influenza A viruses, are currently co-circulating in swine populations in Korea. An outbreak of the highly pathogenic avian influenza H5N1 virus occurred in domestic bird farms in Korea during the winter season of 2003. Pigs can serve as hosts for avian influenza viruses, enabling passage of the virus to other mammals and recombination of mammalian and avian influenza viruses, which are more readily transmissible to humans. This study reports the current seroprevalence of swine H1 and H3 influenza in swine populations in Korea by hemagglutination inhibition (HI) assay. We also investigated whether avian H5 and H9 influenza transmission occurred in pigs from Korea using both the HI and neutralization (NT) tests. 51.2% (380/742) of serum samples tested were positive against the swine H1 virus and 43.7% (324/742) were positive against the swine H3 virus by HI assay. The incidence of seropositivity against both the swine H1 virus and the swine H3 virus was 25.3% (188/742). On the other hand, none of the samples tested showed seropositivity against either the avian H5 virus or the avian H9 virus by the HI and NT tests. Therefore, we report the high current seroprevalence and co-infectivity of swine H1 and H3 influenza viruses in swine populations and the lack of seroepidemiological evidence of avian H5 and H9 influenza transmission to Korean pigs. 相似文献