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猪繁殖—呼吸道综合征病毒（PRRSV）CH—1a株基因型鉴定 总被引：29，自引：4，他引：25
根据猪繁殖－呼吸道综合征病毒（ＰＲＲＳＶ）美洲型毒株和欧洲型毒株的基因组序列设计合成了２对引物,即１００８ＰＳ／１００９ＰＲ和１０１０ＰＬＳ／１０１１ＰＬＲ。我国分离的ＣＨ－１ａ株用这２对引物均能扩增出与预期大小相符的ＲＴ－ＰＣＲ产物,分别与美洲型代表株（ＶＲ－２３３２）的ＲＴ－ＰＣＲ产物大小相当。特异性试验表明,这２对引物均不能扩增其他常见的繁殖障碍相关病毒的ＲＮＡ或ＤＮＡ以及未感染的ＭＡＲＣ－１４５细胞ＲＮＡ。间接免疫荧光试验也证明,ＣＨ－１ａ株与ＰＲＲＳＶ核衣壳蛋白特异性单克隆抗体以及美洲型毒株抗血清均呈阳性反应。因此初步证实ＣＨ－１ａ株为美洲型ＰＲＲＳＶ。 相似文献
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我国猪繁殖和呼吸综合征病毒基因型鉴定 总被引：13，自引：0，他引：13
应用ＲＴ－ＰＣＲ技术,从我国东北和华北地区分离的２个猪繁殖和呼吸综合征病毒（ＰＲＲＳＶ）毒株中扩增获得部分核衣壳蛋白基因。该基因片段长度与美洲型野毒株ＶＲ２３３２ＲＴ－ＰＣＲ扩增片段相同,均为４３３ｂｐ,长于欧洲型Ｌｅｌｙｓｔａｄ毒株扩增片段长度（３９５ｂｐ）。此外,用另１对引物,从这２个分离毒株及ＶＲ２３３２毒株扩增获得部分ＧＰ５蛋白基因,其限制性酶切片段长度多态性分析结果相同或相似。该结果表明,我国不同地区流行的ＰＲＲＳＶ可能属于同一毒株,并且来源于美洲。 相似文献
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本研究在国内首次成功建立了辣根过氧化物酶标记的链霉亲和素－生物系（ＬＳＡＢ）免疫组化染色法检测猪生殖－呼吸道综合征病毒（ＰＲＲＳＶ）抗原。应用ＬＳＡＢ染色技术检测１２头人工感染ＰＲＲＳＶ美洲株（ＡＴＣＣ ＶＲ－２３３２）或国内分离株（Ｂ９６－４，Ｂ９６－５）的ＳＰＦ仔猪组织细胞内的ＰＲＲＳＶ抗原，阳性检出率为１００％。 相似文献
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斑点酶联免疫吸附试验检测猪繁殖与呼吸综合征病毒抗体方法的建立 总被引：6，自引：0，他引：6
首次建立了斑点－酶联免疫吸附试验（Ｄｏｔ－ＥＬＩＳＡ）检测猪繁殖与呼吸综合征病毒（ＰＲＲＳＶ，美洲株）抗体的方法。特异性鉴定表明，ＰＲＲＳＶ不与猪瘟病毒、猪伪狂犬病病毒、猪细小病毒阳性血清反应；与美国进口的ＰＲＲＳＶ抗体ＥＬＩＳＡ诊断试验盒检测结果比较，３５份猪血清的阴、阳性检出总符合率为８２．９％（２９／３５），其中Ｄｏｔ－ＥＬＩＳＡ的阳性检出率略高于进口试剂盒的检出率。 相似文献
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猪生殖—呼吸道综合征病毒CH—1al株ORF2～7序列测定 总被引：2，自引：0，他引：2
新近发现的猪生殖－呼吸道综合征病毒（ＰＲＲＳＶ）是单股ＲＮＡ病毒，属不久前成立的动脉炎病毒科。为了比较从国内分离的ＰＲＲＳＶ与欧美ＰＲＲＳＶ毒株的分子遗传学关系，本文扩增并克隆了ＰＲＲＳＶ ＣＨ－１ａ株ＯＲＦ２￣７，测定了其核苷酸序列。与北美洲型代表株ＶＲ－２３３２遗传距离最近，可能来自同一祖先。 相似文献
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用反转录—聚合酶链反应检测猪繁殖与呼吸综合征病毒的研究 总被引：6，自引：1，他引：5
根据猪繁殖与呼吸综合征病毒美洲型标准毒株的ＯＲＦ６及部分ＯＲＦ７的序列，设计合成了一对引物２６３７４ ＰＳ／２６３７５ＰＲ，用反转录－聚合酶链反应技术对６株ＰＲＲＳＶ北京分离株进行了检测。结果该引物对６株病毒均能扩增出与预期大小相符的ＲＴ－ＰＣＲ产物，并与ＡＴＣＣ ＶＲ－２３３２株扩增产物的大小相当，而欧洲型标准毒株未获扩增片段。 相似文献
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猪繁殖呼吸综合征 (ＰＲＲＳ)是由ＰＲＲＳＶ引起的传染病。 1 997年我们从上海郊县发病猪场的死胎和死仔中分离到ＰＲＲＳＶＳ1毒株 ,经鉴定属美洲型毒株〔1〕。ＰＲＲＳＶ对母猪和仔猪易感 ,感染后在临床症状方面可分二大类 ,一类是母猪表现为繁殖障碍 ;另一类是仔猪和育成猪表现出呼吸道症状。本试验将ＰＲＲＳＶ上海分离株接种易感仔猪 ,以复制出ＰＲＲＳ病症 ,从而进一步确证分离到的Ｓ1毒株属ＰＲＲＳＶ。1 材料和方法1 .1 试验用仔猪 ,从上海远郊一个ＰＲＲＳ阴性猪场购买 ,隔离饲养 ,经检测抗体阴性 ,作为本试验用猪。1 .2 病毒 … 相似文献
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猪繁殖─呼吸道综合症黄光红云南省畜牧兽医学校（６５０２１２）猪繁殖─呼吸道综合症（ＰＲＲＳ）是一种新发生的猪传染病。ＰＲＲＳ是由猪繁殖一呼吸道综合症病毒（ＰＲＲＳＶ）或称累利斯塔德病毒（ＬｅｌｙｓｔａｄＶｉｒｕｓ，ＬＶ）引起的。其病的特征是繁殖母猪的... 相似文献
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检测猪繁殖与呼吸综合征病毒抗体ELISA方法的建立 总被引：19，自引：0，他引：19
本试验通过差速离心法和非线性蔗糖密度梯度离心法纯化猪繁殖与呼吸综合征病毒（ＰＲＲＳＶ）。经三氯－三氟乙烷处理后,作为ＥＬＩＳＡ试验的标准抗原,并建立了检测ＰＲＲＳＶ抗体ＥＬＩＳＡ方法。该方法与ＩＦＡ、ＩＰＭＡ和国外同类商品化试剂盒进行了对比试验,表明该方法具有敏感性高、特异性好的特点,是一种快速准确检测ＰＲＲＳＶ抗体的方法。间接ＥＬＩＳＡ方法在敏感性上要优于ＩＦＡ和ＩＰＭＡ》 相似文献
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为快速准确区分PRRS流行毒株与减毒活疫苗TJM F92株以及对流行毒株进行定量检测,按GenBank中发表的美洲型PRRSV SX 1分离株、弱毒疫苗株NSP2基因缺失部位的不同设计了一对特异性引物,通过RT PCR和体外转录的方法,构建了体外转录RNA作为标准品,并对反应条件和反应体系进行优化,旨在建立一种敏感性高、特异性强、重复性和稳定性良好的qPCR鉴别方法。结果表明,该方法最低能检测出1.0×101拷贝/μL的模板,敏感性比常规PCR高100倍;应用该方法对218份临床样本进行鉴别与定量,qPCR检出率较常规RT PCR高12.9个百分点。研究表明,qPCR鉴别方法的建立实现了对PRRS流行毒株与疫苗毒株的快速区分以及对流行毒株的定量检测,为PRRS的快速诊断提供了依据。 相似文献
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van der Linden IF van der Linde-Bril EM Voermans JJ van Rijn PA Pol JM Martin R Steverink PJ 《Veterinary microbiology》2003,97(1-2):45-54
The current study was performed to determine if porcine reproductive and respiratory syndrome virus (PRRSV) could be transmitted to pigs by feeding muscle tissue obtained from recently infected pigs. Muscle obtained from pigs infected with either a European strain (EU donor pigs) or American strain (US donor pigs) of PRRSV was fed to PRRSV-free receiver pigs. The donor pigs were slaughtered 11 days post-infection (dpi). PRRSV was detected by conventional virus isolation in muscle at 11 dpi from 7 of 12 EU donor pigs and 5 of 12 US donor pigs. In contrast to conventional virus isolation, all muscle samples from infected pigs were positive for viral nucleic acid by PCR, except for muscle from one animal infected with the American strain of PRRSV. Five hundred grams of raw semimembranosus muscle from each of the donor pigs was fed over a 2 days period (250 g per day) to each of two receiver pigs (48 receiver pigs). The receiver pigs were housed separately in five groups. One of the five groups was fed muscle obtained from US donor pigs that was also spiked with the American strain of PRRSV. Sentinel pigs were placed in-contact with the group of receiver pigs fed spiked muscle. All receiver pigs became viraemic by 6 days post-feeding (dpf). There was evidence of horizontal transmission with sentinel pigs, in-contact with receiver pigs, becoming viraemic. The study demonstrates that PRRSV could be infectious through the oral route via the feeding of meat obtained from recently infected pigs. 相似文献
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In Denmark, a porcine reproductive and respiratory syndrome virus (PRRSV) control programme, comprising vaccination of seropositive herds with a live American type PRRSV vaccine, was started in 1996. In several of these herds, spread of vaccine virus from vaccinated 3-18 week old pigs to non-vaccinated sows was demonstrated by the isolation of vaccine virus from fetuses and stillborn piglets. Surprisingly, sows infected with the American type vaccine strain consistently exhibited significantly stronger serological responses towards European type PRRSV than American type PRRSV. In order to elucidate whether the unexpectedly strong serological reaction towards European-type PRRSV in American type PRRSV infected sows was due to a booster reaction, or reactivation of an unrecognized, latent infection in the sows with European type PRRSV, a challenge study with the vaccine was carried out. In this study, the stronger serological response towards European type PRRSV than towards American type PRRSV was reproduced, and reactivation of the previous natural infection with European PRRSV could neither be demonstrated by virus isolation nor by RT-PCR. So, the increase in antibody titers towards European PRRSV in previously European PRRSV infected pigs after challenge with the vaccine strain seems to be the result of a boosting effect on the immune system, induced by the heterologous vaccine PRRSV strain. 相似文献
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Rapid detection of porcine reproductive and respiratory syndrome viral nucleic acid in blood using a fluorimeter based PCR method 总被引：4，自引：0，他引：4
Spagnuolo-Weaver M Walker IW Campbell ST McNeilly F Adair BM Allan GM 《Veterinary microbiology》2000,76(1):15-23
Porcine reproductive and respiratory syndrome virus (PRRSV) is an Arterivirus recognised world wide as an important cause of reproductive failure and pneumonia in pigs. American and European strains of PRRSV, differentiated antigenically and genomically, have been reported. PRRSV infections are currently diagnosed using serology, virus isolation and/or immunocytochemistry. In order to overcome various drawbacks associated with these techniques, conventional, block-based RT-PCR methods for the detection of PRRSV nucleic acid in clinical samples have been described. These methods require gel electrophoresis for analysis of PCR products and present high risk of DNA carry-over contamination between the samples tested. We describe the detection of PRRSV RNA in serum samples and in blood impregnated filter disks (FDs), obtained from experimentally inoculated pigs, using a closed-tube, fluorimeter-based PCR assay. The assay eliminates the use of gel electrophoresis, and is as sensitive and specific as the conventional block-based PCR assay, detecting positive samples as early as 1 day post-inoculation. We also report a rapid fluorimeter based PCR method for differentiating American and European strains of PRRSV. 相似文献
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对PRRSV快速分型的多重RT-PCR方法的建立与应用 总被引：4，自引：2，他引：2
根据猪繁殖与呼吸综合征病毒(PRRSV)欧洲株、美洲经典株与高致病性Nsp2变异株的Nsp2基因间的差异,设计3对特异性扩增引物,建立快速检测PRRSV欧洲株、美洲经典株和高致病性Nsp2变异株的多重RT-PCR方法。利用PRRSV欧洲株、经典株和高致病性Nsp2变异株及其他相关病毒株进行敏感度和特异性试验。结果显示,所建立的多重RT-PCR对欧洲株、美洲经典株与高致病性Nsp2变异株的RNA最小检出量分别为0.050 2、0.055 4、0.056 4ng,与CSFV、TGEV、JEV、SIV的核酸均无交叉反应。用该方法检测病料76份,结果PRRSV欧洲株、美洲经典株和高致病性Nsp2变异株的阳性率分别为0、7.89%和53.9%,未发现3型病毒间有混合感染的情况。检测结果与单一RT-PCR及RT-PCR检测试剂盒检测结果一致,高于病毒分离。 相似文献