共查询到19条相似文献，搜索用时 408 毫秒
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用PCR检测长沙市猪肺炎支原体和猪鼻支原体的感染情况 总被引：1，自引：0，他引：1
为对长沙市猪支原体肺炎病原流行病学进行初步调查,用PCR方法对从湖南省长沙市5个县（市、区）205头猪中收集的326份病料（205份猪肺和121份猪鼻拭子）分别进行了猪肺炎支原体和猪鼻支原体检测,并对部分基因进行了序列分析。结果发现,猪肺炎支原体检出率为24.4%（50/205）,猪鼻支原体检出率为22.3%（27/121）;长沙市每个县（市、区）的生猪中均存在这两种支原体,其中长沙县生猪阳性率最高（42.4%和31.5%）;序列分析表明,检测出的基因序列与国内外相关序列有97%～100%的同源性。本试验证实了猪肺炎支原体和猪鼻支原体在长沙市猪场的存在。 相似文献
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《养猪》2016,(1)
用PCR方法对从湖南省长沙市515头屠宰猪收集的1 204份样品(120份猪鼻拭子、254份猪支气管拭子、515份猪支气管肺泡灌洗液和235份猪肺、80份血液)进行了猪肺炎支原体和猪鼻支原体检测,并对部分PCR产物进行序列分析。结果发现,猪肺炎支原体在猪支气管灌洗液中检出率最高(23.3%,120/515),猪鼻拭子和血液中检出率最低(0);猪鼻支原体在猪鼻拭子中检出率最高(24.3%,125/515),在猪支气管拭子中检出率最低(2.5%,3/120)。序列分析表明:试验的PCR产物与国内已出版的相关序列有97%~100%的同源性。试验探明了猪肺炎支原体和猪鼻支原体在猪呼吸道与血液中的分布情况,为这两种支原体的分离鉴定提供了依据。 相似文献
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猪传染性胸膜肺炎放线杆菌PCR检测方法的建立及应用 总被引：1，自引：0，他引：1
猪传染性胸膜肺炎是由胸膜肺炎放线杆菌引起猪的一种高度接触性呼吸道传染病,该病可给养猪业造成巨大的经济损失。为有效控制和确诊该病,根据报道的猪胸膜肺炎放线杆菌APXIV毒株的基因序列,合成了2对可扩增长度分别为442 bp和378 bp的特异引物,建立检测胸膜肺炎放线杆菌的巢式PCR方法。利用合成的引物在扩增猪肺疫巴氏杆菌、猪链球菌、副猪嗜血杆菌和大肠杆菌等细菌DNA时,结果均为阴性。用引物检测猪胸膜肺炎放线杆菌的标准菌株可扩增出442 bp和378 bp的特异性条带。表明运用PCR法检测猪胸膜肺炎放线杆菌的特异性和灵敏性均较高,可作为猪传染性胸膜肺炎的快速诊断和流行病学调查的手段。 相似文献
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猪传染性胸膜肺炎放线杆菌PCR检测试剂盒的研制 总被引：1，自引：1，他引：0
根据GenBank中猪传染性胸膜肺炎放线杆菌apxⅣA基因序列,设计了1对引物,通过对PCR反应条件进行优化,研制了检测猪传染性胸膜肺炎放线杆菌的PCR试剂盒。该试剂盒扩增的阳性条带为600 bp,特异性与敏感性结果显示,该PCR检测试剂盒的最低核酸检测量为50 CFU/mL,而对金黄色葡萄球菌、链球菌、多杀性巴氏杆菌、鼠伤寒沙门氏菌、副猪嗜血杆菌、大肠杆菌的扩增结果均为阴性。-20℃至少可保存12个月,且重复性良好。应用该PCR试剂盒对41份临床样本进行了检测,其PCR检测结果与细菌学检测结果相一致。结果表明,猪传染性胸膜肺炎放线杆菌PCR检测试剂盒能够对APP临床样品进行快捷、灵敏、准确的检测。 相似文献
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《黑龙江畜牧兽医》2016,(20)
为了解天津部分猪场副猪嗜血杆菌病(HPs)和猪传染性胸膜肺炎(PCP)的流行情况,利用间接血凝试验对采自9个规模猪场的178份猪血清样品进行副猪嗜血杆菌抗体检测,对采自8个规模猪场的166份猪血清样品进行猪传染性胸膜肺炎放线杆菌抗体检测。结果表明:副猪嗜血杆菌阳性率为18.54%(33/178),其中断奶仔猪和保育阶段猪阳性率为23.58%(25/106);猪传染性胸膜肺炎的阳性率为21.08%(35/166),其中断奶仔猪阳性率为20.93%(9/43),育肥猪阳性率为27.66%(13/47),种猪阳性率为21.62%(8/37)。说明天津地区部分猪场的副猪嗜血杆菌病和猪传染性胸膜肺炎呈地方性流行。 相似文献
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河南省周口地区猪传染性胸膜肺炎的血清学调查 总被引：1，自引：0，他引：1
猪传染性胸膜肺炎（Porcine Cntagious Pleuropneumonia）是由胸膜肺炎放线杆菌（ Actinobacillus Pleuropneumoniae,APP）引起的一种猪的呼吸道传染病,能引起猪急性纤维素性胸膜肺炎或慢性局灶性坏死性肺炎。该病是世界范围内的多发病,近几年发病率呈逐年上升趋势。河南省周口地区具有较大的养猪规模,为及时掌握周口地区猪传染性胸膜肺炎的流行动态,我们于2007年10月份对周口地区的8个县（市）部分规模化养猪场随机采取544份猪血清进行了血清学检测,现将结果报告如下。 相似文献
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猪传染性胸膜肺炎（In- fections pleuropneumonia of swine,IPPS）是由胸膜肺炎放线杆菌（Actionbacillus pJeuropneumoniae,App）引起的一种猪的呼吸道传染病,各种年龄猪均可感染,2-5月龄猪最易发生,具有高度传染性,常与巴氏杆菌混合感染。 相似文献
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Fablet C Marois C Dorenlor V Eono F Eveno E Jolly JP Le Devendec L Kobisch M Madec F Rose N 《Research in veterinary science》2012,93(2):627-630
Relationships between macroscopic lesions and Polymerase Chain Reaction (PCR) detection of Mycoplasma hyopneumoniae (Mhp), Pasteurella multocida (Pm), Actinobacillus pleuropneumoniae (App), Haemophilus parasuis (Hps) and Streptococcus suis (Ssuis) of the lungs of 3731 slaughter pigs from 125 herds were assessed in France. Pneumonia and pleuritis were the most frequent lesions (69.3% and 15% of the lungs, respectively). Mhp, Pm, App, Ssuis and Hps were detected in 69.3%, 36.9%, 20.7%, 6.4% and 0.99% of the lungs, respectively. Mhp and Pm were associated with pneumonia at both the pig and herd levels. Pleuritis was not associated with any pathogen at the pig level, but was associated with a high percentage of pigs PCR-positive for App at the herd level. Measures focused on control of Mhp, Pm and App should significantly reduce the occurrence of both pneumonia and pleuritis. 相似文献
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Szczotka A Stadejek T Zmudzki J Nowak A Osiński Z Pejsak Z 《Polish journal of veterinary sciences》2011,14(4):531-538
The aim of the present study was to develop an immunohistochemical method (IHC) for detection of Lawsonia intracellularis (L. intracellularis) in formalin-fixed, paraffin embedded sections of intestines from pigs and to implement this method in differential diagnosis of swine diseases with diarrhea in postweaning pigs. The study was conducted on 165 sections of intestines (ileum, caecum and colon) collected from 76 pigs, representing 42 Polish pig farms. The animals included in the analysis suffered from diarrhea, with bloody or grey to brown feces, and were suspected of porcine proliferative enteropathy (PPE). Sections of intestines were analyzed for the presence of L. intracellularis by polymerase chain reaction (PCR) and IHC. Among 165 intestinal samples from pigs with diarrhea, L. intracellularis DNA was detected by PCR in 33 (20.0%) samples. In this group, 30 samples (18.2% of all the samples tested) were also found positive in IHC, while only 3 (1.8%) were IHC-negative. One hundred thirty-two (80.0%) samples were negative in both tests. The PCR- and IHC-positive samples originated from 11 pigs, 4- to 20-week old, from 8 farms. L. intracellularis antigen was visualized by IHC mostly in intestinal crypts and/or in mononuclear cells of the lamina propria). The positive signal in epithelial cells was observed close to the luminal borders, creating typical specifically stained rims around the crypt lumina. The results of the present study further confirm the usefulness of IHC in the detection of L. intracellularis antigen in the intestinal tissues. 相似文献
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A PCR assay used to study aerosol transmission of Actinobacillus pleuropneumoniae from samples of live pigs under experimental conditions 总被引：2，自引：0，他引：2
Savoye C Jobert JL Berthelot-Hérault F Keribin AM Cariolet R Morvan H Madec F Kobisch M 《Veterinary microbiology》2000,73(4):337-347
The study describes a polymerase chain reaction (PCR) assay for the detection of Actinobacillus pleuropneumoniae. The test is based on the amplification of the omlA gene coding for an outer membrane protein of A. pleuropneumoniae. To test the specificity of the reaction, 19 other bacterial species related to A. pleuropneumoniae or isolated from pigs were assayed. They were all found negative in the PCR assay. The detection threshold of the test was 10(2) A. pleuropneumoniae CFU/assay. The test was then applied to the detection of A. pleuropneumoniae from tonsillar biopsies and tracheobronchial lavage fluids of pigs without a culture step. The detection of A. pleuropneumoniae in these samples was performed by PCR, by conventional culture and by bacteriology with immunomagnetic beads. The number of samples that were found positive by PCR was almost three times higher than the number of samples from which A. pleuropneumoniae was isolated by both bacteriological techniques. The detection of A. pleuropneumoniae in these samples allowed us to demonstrate its aerosol transmission to pigs under experimental conditions. The trial involved 18 specific pathogen free pigs. Six pigs, infected with A. pleuropneumoniae, were located in a unit A, together with four non-infected animals (contact pigs). Eight non-infected pigs (reporter pigs) were located in a unit B, adjacent to A. We detected A. pleuropneumoniae in samples from infected animals but also from 'contact' (unit A) and 'reporter' (unit B) pigs. The results of this study show that the simple preparation of the samples followed by the PCR assay may be a useful tool for epidemiological studies. 相似文献
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Arnold T Scholz HC Marg H Rösler U Hensel A 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2004,51(10):459-463
This study evaluated the suitability of invA gene amplification by PCR as an effective means of detecting Salmonella species in pigs experimentally infected with S. Typhimurium DT104. A controlled infection study using 24 pigs was performed in order to compare efficacy, precision and detection rates of the invA-based PCR method originally described by Rahn, K. De Grandis, S.A., Clarke, R.C., McEwan, S.A., Galan, J.E., Ginocchio, C., Curtiss, R. 3rd, C.L. Gyles, (Mol. Cell. Probes 1992; 6: 271-279) as a new in-house invA-based PCR method for the specific detection of Salmonella spp. in pork and different tissue samples of slaughter pigs. Finally, PCR results were compared with culture detection rates obtained by isolation procedures following the ISO 6579:2000, the 'gold standard'. After slaughtering, 14 different tissue samples of each pig were investigated to verify the usefulness of the two invA-based PCR methods in different matrices of slaughter pigs. The results demonstrate that the application of the widely used invA-based primer pair (139 + 141) may result in questionable products if samples gained from selective enrichment in the Rappaport-Vassiliadis medium were investigated. These questionable products can lead to false-positive results, if no additional hybridization procedure is attached or if unspecialized persons use this method in routine laboratory practice. The newly developed in-house PCR method used is based on the 3'-prime region of invA, especially designed and harmonized for the detection of Salmonella in different matrices of slaughtered pigs after bacterial enriched broth culture. In this study, this PCR revealed no questionable products and, furthermore, the specificity of the amplificate could be tested by means of the restriction enzyme NdeI. In comparison with the culture detection procedure, the new PCR method has a sensitivity of 100% and a specificity of 96%. Thus, this method might be used as a meaningful tool in eliminating Salmonella-positive carcasses at slaughterhouse level and thus, keeping them out of the food chain. 相似文献
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Jung K Ha Y Ha SK Kim J Choi C Park HK Kim SH Chae C 《Veterinary journal (London, England : 1997)》2006,171(1):166-168
The identification of porcine circovirus type 2 (PCV2) was studied in fresh intestinal tissues by polymerase chain reaction (PCR) and in formalin-fixed, paraffin-wax-embedded intestinal tissues by in situ hybridisation. The tissues came from pigs naturally infected with porcine epidemic diarrhoea virus (PEDV). A total of 35 (32.7%) of 107 small intestinal samples from pigs naturally infected with PEDV were found to be positive using PCR. Positive signals for PCV2 were detected in 32 (29.9%) of 107 small intestinal samples from pigs naturally infected with PEDV by in situ hybridisation. The distribution of positive cells in the jejunum and ileum was multifocal or patchy. Distinct positive labelling was found throughout the lamina propria in the small intestines. The results of this study indicate that PCV2 is highly prevalent in pigs naturally infected with PEDV. 相似文献
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The goal of this study was to estimate the accuracy of the invA-gene-based polymerase chain reaction (PCR) and a culture technique based on pre-enrichment with buffered peptone water, three selective enrichment media (selenite, tetrathionate and Rappaport-Vassiliadis broths) and four selective, solid media (Xylose-Lysine-Tergitol-4, Salmonella/Shigella, Hekton-Enteric and MacConkey), for the detection of Salmonella organisms from caecal samples from slaughter pigs. For this purpose a latent-class (Bayesian) approach was used. Two hundred and three slaughtered pigs were used after grouping them into two groups of 96 and 107 animals. Sensitivity (Se) was estimated to be 56% (95% probability interval 40, 76) for culture and 91% (81, 97) for PCR. The specificity (Sp) of the PCR was 88% (80, 95) while the Sp of the culture had been considered 100% in the statistical analysis as all culture-positive samples were confirmed by serotyping. PCR Se was not affected by the Salmonella serotypes present in the samples analysed. Accordingly, a minimum of 25.5% of the pigs was estimated to harbour Salmonella organisms in their faeces. It was concluded that bacteriology on caecal samples alone was a poor diagnostic method, and that the PCR method could be considered a cost-effective alternative to culture in Salmonella monitoring programmes. However, given the moderate Sp of this molecular technique, PCR-positive samples should be further confirmed through bacteriology. 相似文献
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Loop-mediated isothermal amplification (LAMP) is a simple method that can amplify DNA with high specificity, sensitivity, and rapidity. In this study, we compared the performance of LAMP and real-time PCR assays for diagnosis of toxoplasmosis. We designed a real-time PCR assay targeting a 529 bp element repeated 200-300 times in the Toxoplasma gondii genome. The detection limits of the LAMP and real-time PCR assays were 10 fg/μL and 1 fg/μL of T. gondii DNA, respectively. Conventional PCR, LAMP, and real-time PCR methods were applied to detect T. gondii DNA in blood samples from 284 pigs and 292 sheep. Positive results were obtained with 0.4%, 3.2%, and 4.2% of the pig samples and 3.8%, 17.1%, and 17.8% of the sheep samples with conventional PCR, LAMP, and real-time PCR analyses, respectively. The real-time PCR assay provided the most sensitive diagnosis of toxoplasmosis, but the LAMP assay has potential as an alternative tool for detection of T. gondii in the field. 相似文献
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江苏地区猪圆环病毒2型流行病学调查 总被引：2，自引：0，他引：2
应用复合PCR方法,对江苏地区的162份可疑病料进行了检测,结果72份病料为PCV2阳性,17份为PCV1阳性,其中7份同时感染PCV1和PCV2。从发病时间看,以9~11月发病最多,阳性率高达54.67%。样品较多的保育猪和育肥猪的阳性率分别是26.03%和68.12%。通过流行病学调查,表明该病已在江苏地区呈流行趋势。同时对18头PCV2阳性猪病料的心、肝、脾、肺、肾、脑、腹股沟淋巴结、肠系膜淋巴结、扁桃体进行PCV2检测,结果有7头PCV2阳性猪的各个脏器都能检测出PCV2;在这些脏器中,肺、腹股沟淋巴结、扁桃体的检出率都为100%,较其他受检脏器更适合作为PCV2的检测样品。 相似文献
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根据GenBank中已发表的猪细小病毒(porcine parvovirus,PPV)、猪伪狂犬病病毒(pseudorabies virus,PRV)和猪圆环病毒2型 (porcine circovirus type 2,PCV2)基因序列,对各病毒基因区进行同源性分析,确定PPV 的VP2、PRV的 gD、和PCV2的ORF2基因为各病毒的诊断靶序列,设计特异性引物,在建立各病毒单项PCR技术的基础上,优化多重PCR反应条件,建立了3种病毒的多重PCR技术,可同时扩增PPV 313 bp、 PRV 217 bp和PCV2 447 bp的特异性片段。用多重PCR技术与单项PCR技术对比检测试验证明两者的符合率为100％,表明建立的多重PCR检测方法,具有特异、快速、准确的特点,可同时鉴别诊断这3种病毒。从10个发病猪场和门诊病例的病猪采集的211份样品,用建立的多重PCR检测方法,检出PPV阳性42份,阳性率为19.91%;PRV阳性26份,阳性率为12.32%;PCV2阳性56份,阳性率为26.54%;2种以上病毒混合感染25份,混合感染阳性率为11.85%。检测结果表明,山西省猪群已感染这3种疫病。 相似文献