不同方法检测鸡白痢沙门氏菌抗体消长规律的比较研究

为比较不同方法检测鸡白痢沙门氏菌抗体消长的规律,以便为种鸡场净化提供科学指导,本研究以鸡白痢沙门氏菌活菌和灭活免疫原分别接种SPF鸡,采用平板凝集、微量凝集和ELISA试验定期检测血清中的特异性抗体,并以Kappa检验判定不同检测方法之间的一致性程度。结果显示,平板凝集试验在接种后检出抗体阳转的时间早于ELISA,但ELISA检出抗体阳性的持续时间更长,且更符合抗体消长规律;3种检测方法的结果之间仅具有微弱一致性(Kappa系数为0.002~0.295)。本研究结果表明,不同鸡白痢沙门氏菌抗体检测方法之间存在较大差异,但从总体来看,ELISA无假阳性干扰,检出抗体的持续时间较长,更符合抗体消长规律,在单一鸡白痢沙门氏菌感染的情况下,更有利于种鸡场对该病进行净化。     

鸡白痢/禽伤寒沙门菌抗体检测试剂的比较与评估

对市面上现有鸡白痢/禽伤寒沙门菌抗体平板凝集检测试剂及其不同批次,微量凝集检测试剂和ELISA共4种检测试剂进行比较与评估。用4种试剂以及试剂A的不同批次检测来自11个种鸡场的1 650份血清,计算不同试剂检测种鸡场的抗体阳性率,2种试剂之间的阳性符合率、阴性符合率和总符合率,并以Kappa检验判断不同检测方法及其试剂之间的一致性程度。平板凝集试剂A的不同批次间以及平板凝集试剂A和B之间具有高度一致性(Kappa系数=0.714~0.746),但阳性符合率不足80%。平板凝集试剂A、微量凝集试剂C、ELISA试剂D之间仅具有中度或弱一致性(Kappa系数=0.392~0.542)。不同鸡白痢/禽伤寒沙门菌抗体检测试剂间存在差异,平板凝集试剂不同批次间也存在差异,提示试剂生产厂家应加强质量控制,同时研发敏感性、特异性、稳定性更好的替代试剂,而种鸡场在进行抗体监测时应固定使用1种检测试剂,避免影响检测结果的准确性和连续性。     

琼脂扩散法排除平板凝集法中的假阳性鸡白痢

<正> 一、全血平板凝集不但能检出鸡白痢特异性抗体而且也能检出非异性抗体(类属抗体)。上表说明鸡白痢全血平板凝集阳性血清不但能凝集白痢沙门氏菌,而且也能凝集大肠杆菌、绿脓杆菌、变形杆菌、葡萄球菌。因为这些细菌具有和鸡白痢沙门氏菌抗原中相同或相近似的抗原,白痢鸡血清中有特     

竹丝鸡和PSF鸡人工感染鸡白痢沙门菌的抗体消长规律

本研究通过人工感染鸡白痢沙门菌到竹丝鸡和SPF鸡,采用血清平板凝集的方法检测鸡白痢抗体,用于确定鸡白痢抗体的消长规律。结果表明:不同品种间鸡性成熟时间具有差异,鸡白痢抗体高峰出现时间也不完全相同。在不同感染方式下鸡白痢抗体消长规律具有差异,腹腔注射组抗体产生最快,滴鼻组次之,同居感染组抗体产生最慢。     

全血与血清平板凝集试验检测鸡白痢的对比

对北京油鸡两个品系216只母鸡,在17~49周期间每隔8周同时用全血和血清平板凝集试验进行鸡白痢检测对比分析。结果发现,血清检出阳性率高于全血检出阳性率,且血清检出阳性个体包含全血检出阳性个体;同时,检测结果也表明,33周龄鸡白痢阳性率达到最高,此后阳性率下降。因此,在鸡白痢净化过程中采用血清平板凝集法比全血平板凝集法检测更为敏感,在25~33周对北京油鸡群体增加一次鸡白痢净化更有助于加快净化进程。本研究可为其他地方品种种鸡鸡白痢检测工作提供参考。     

人工感染鸡白痢菌抗体消长规律的试验研究

为辽宁省鸡白痢净化,提供最佳检验时机。我站历经一年的时间,用连续6年净化鸡场,所孵化的鸡雏140只。应用平板凝集实验,琼脂扩散试验等检验方法。模拟规模饲养场的饲养条件,人工感染鸡白痢菌,检测抗体消长规律。最后通过97只鸡的检测结果证明;平板凝集抗体在49～52日龄为抗体阳性的高峰时间,凝集抗体的高峰时间可维持13～19d。116日龄后,多数平板凝集抗体转为阴性,215日龄后,没有新的抗体阳性鸡出现。琼脂扩散沉淀抗体在26～58日龄和123～251日龄抗体阳性率最高。     

鸡白痢检疫应注意的几个问题

鸡白痢检疫应注意的几个问题崔统一（济南市畜禽良种繁育中心２５０１００）当前，种鸡场鸡白痢的净化工作，主要是采用鸡白痢染色平板凝集抗原对鸡群逐只进行全血平板检疫，淘汰阳性反应鸡的方法。通过这种方法逐步消除鸡白痢沙门氏菌感染鸡，最终达到净化之目的。作者根...     

鸡白痢检疫值得注意的几个问题

鸡白痢检疫值得注意的几个问题康凯（中国兽药监察所，北京１０００８１）我国种鸡场鸡白痢的净化工作，主要是采用鸡白痢染色平板凝集抗原对鸡群逐只进行全血平板检疫，淘汰阳性反应鸡的方法。通过这种方法逐步消除鸡白痢沙门氏菌感染鸡，最终达到净化之目的。经几年的净...     

种鸡场沙门菌血清学与病原学监测结果比较与分析

为了解种鸡场鸡白痢抗体与携带沙门菌之间的相关性,以便选择合适的检测方法用于种禽场疫病净化,本研究在全国种鸡养殖重点区域内选择12个种鸡场,随机采集血清、泄殖腔拭子和盲肠样品进行鸡白痢抗体检测和沙门菌的分离鉴定。结果显示,检出抗体阳性的种鸡场共9个,而分离到沙门菌的种鸡场仅有5个;同时,检测发现部分种禽场抗体检测结果为阴性但病原检测为阳性。上述结果表明,抗体阳性率与病原阳性率并不一致,提示种鸡场在进行相关疫病净化时,应综合考虑血清学和病原学检测结果,防止出现因漏检而造成全群污染的情况发生。     

鸡白痢沙门氏菌感染情况的调查

鸡白痢是由鸡白痢沙门氏菌引起的一种败血性细菌性疾病 ,它能垂直传递和水平传播。为了控制本病 ,对种鸡群进行净化是最根本的措施。鸡白痢净化的主要方法是检测淘汰阳性种鸡和防止再次感染。检测方法主要是采用现场全血平板凝集试验。为了最大限度检出和淘汰阳性鸡 ,连续进行了 4次检测。1　材料和方法鸡白痢平板凝集抗原 :由中国兽药检察所生产的鸡白痢鸡伤寒多价染色抗原。鸡白痢阳性和阴性对照血清 :由中国兽药检察所提供。检测方法和判断标准 :全血平板凝集试验。检测时机 :1 6周龄开始 ,全群普检 ,淘汰阳性鸡 ,如阳性率大于 0 .2 % ,…     

Improvements required for the detection of Salmonella Pullorum and Gallinarum

Detection of the specific Salmonella serovar Gallinarum, which is divided into the biovars Pullorum and Gallinarum, is compulsory under the national hygienic and sanitary control regulations of France for breeding flocks whose offspring are exported. Our aim was to examine the suitability of bacteriologic and serologic methods routinely used in France to screen serum samples and organs for S. Gallinarum. Since bacteriologic reference techniques are designed to isolate the commonly occurring non-typhoid serovars, such as S. Typhimurium, S. Enteritidis, and others that cause outbreaks of foodborne illness, they may not be particularly suitable for detecting S. Pullorum and S. Gallinarum. This hypothesis was confirmed by the inoculation of 10-wk-old chickens and 1-d-old chicks with various strains of S. Pullorum and S. Gallinarum. The most reliable enrichment media were selenite cystine and Rappaport-Vassiliadis broths. Moreover, on the usual plating media, colonies were small, grew more slowly than the common serovars (in 48 h instead of 24 h), and had an unusual appearance. Since the rapid slide agglutination (RSA) test is based only on antigens from standard and variant strains of S. Pullorum, it may not readily detect S. Gallinarum. In our study, it detected infection in all 10-wk-old chickens inoculated with S. Pullorum strains but did not detect any antibodies against S. Gallinarum. Therefore, S. Gallinarum antigens must be added to the S. Pullorum antigens used in the RSA test in order to detect antibodies produced by birds infected with either biovar.     

1例鸡白痢沙门氏菌的分离鉴定与耐药性分析

为查明河南省新乡市某规模化蛋鸡场疑似沙门氏菌感染病例的病原，进而制定合理的治疗方案，本研究无菌采集疑似病雏鸡肠管样品，用革兰氏染色、培养特性观察、生化试验、PCR方法对其进行确诊，并进行病原回归试验，采用药敏纸片琼脂扩散法（K-B法）分析分离菌对19种常见抗生素的耐药性。结果显示，本试验成功分离了一株革兰氏阴性短杆菌，该分离菌符合鸡白痢沙门氏菌的培养特性和生化特性；PCR成功扩出invA基因，通过与GenBank数据库比对分析确定该细菌为鸡白痢沙门氏菌；用分离细菌株感染SPF鸡能复制出与自然感染一致的病例，说明鸡白痢沙门氏菌是造成本养鸡场雏鸡发病的主要病原；分离株具有较强的致病性和多重耐药性，对阿米卡星、苯唑青霉素、青霉素耐药，对复达欣、氨苄青霉素、头孢曲松等药物敏感，将敏感抗生素用于临床治疗效果明显。结合以上结果，本研究提出该病的具体防治措施，并取得了较好的效果，为鸡白痢沙门氏菌的分离鉴定及防治提供了参考，对鸡白痢沙门氏菌病病原的早期诊断和治疗具有重要临床意义。     

3种猪伪狂犬病gB抗体ELISA检测试剂盒的比较

采用3种猪伪狂犬病gB抗体ELISA检测试剂盒检测121份血清样品,对检测结果进行Kappa一致性检验。结果显示:3种gB抗体检测试剂盒都具有很好的重复性,两两之间检测结果的符合率达80%以上,Kappa值0.6,表明3种检测试剂盒具有较好的重复性和一致性。     

Evaluation of solid-phase chemiluminescent enzyme immunoassay, enzyme-linked immunosorbent assay, and latex agglutination tests for screening toxoplasma IgG in samples obtained from cats and pigs.

Serum samples from cats and pigs were analyzed by the solid-phase chemiluminescent enzyme immunoassay (SPCEI), enzyme-linked immunosorbent assay (ELISA), and indirect latex agglutination (ILA) methods. The SPCEI and ILA methods accurately analyzed Toxoplasma IgG (T-IgG) in both clinical and spiked samples from pigs and cats. The ELISA method accurately analyzed T-IgG in spiked samples from cats and pigs or clinical samples from pigs, but it did not accurately analyze T-IgG in clinical samples from cats. The antibody used in the ELISA kit did not cross-react with cat T-IgG. The SPCEI method that uses a stand-alone automated analyzer provided quantitative analysis, whereas the ELISA and ILA methods provided qualitative or, at best, semiquantitative analysis of T-IgG. The SPCEI and ELISA methods were rapid (60-90 minutes for 30 samples), whereas the ILA method required 13-15 hours for 30 samples. Although the three methods accurately distinguished positive from negative samples, the ILA method yielded many weakly positive results that were not confirmed by either the ELISA or SPCEI method. Thus, the indirect agglutination tests may give nonspecific responses at lower T-IgG concentrations.     

鸡白痢沙门氏菌病原的分离鉴定及耐药性分析

为鉴定新疆某些地区鸡白痢沙门氏菌主要流行株的流行现状和耐药情况,本研究采用平板凝集试验方法对有发病史的种鸡场进行现场检测,被检的750只鸡中,发现78例疑似鸡白痢沙门氏菌感染,运用沙门氏菌鉴别培养基、组织病理学观察和PCR方法进行确诊,并进行药敏试验。结果显示,疑似的78例鸡白痢沙门氏菌中有69例符合鸡白痢沙门氏菌的生化特性和病理学特点,PCR成功扩增出invA基因,分离率为9.2%,血清型诊断为D1型;其中分离菌对万古霉素和新霉素耐药性最高,耐药率分别为100.0%和81.2%,而对头孢曲松、克拉维酸和多黏菌素的敏感率达60.0%以上。本研究为养殖户和兽医工作者在净化鸡白痢沙门氏菌及临床用药方面提供参考。     

Isolation,Identification and Analysis of Drug Resistance of Salmonella Pullorum

In order to investigate the epidemic situation and drug resistance of major epidemic Salmonella Pullorum in some areas of Xinjiang, 750 chickens with disease history were detected using the methods of plate agglutination, and a total of 78 cases were diagnosed infection with Salmonella Pullorumin by using Salmonella identification medium, histopathological observation, PCR and drug sensitivity test.The results showed that 69 of 78 cases suspect Salmonella Pullorum were isolated, and prove the biochemical characteristics and pathologic characteristics of Salmonella Pullorum, the invA gene was amplified successfully by PCR,the isolation rate was 9.2% and the serotype was diagnosed as D1. The isolates were the most resistant to vancomycin and neomycin, and Salmonella Pullorum resistant rate was 100.0% and 81.2%, respectively. Drug sensitive test results showed that the sensitive rate of the bacteria were more than 60.0% to ceftriaxone,clavulanate and polymyxin. This study provided reference for farmers and veterinary workers, which indicated it could be used for the purification of Salmonella Pullorum and clinical medication.     

Relationship of the enzyme-linked immunosorbent assay to indirect immunofluorescent antibody test for the detection of so-called chicken anemia agent antibodies in serum from broiler breeders.

Serum samples collected from breeder chickens ranging in age from 1 day to 55 weeks were tested for CAA antibodies by enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent antibody (IFA) test. The relationship of ELISA to IFA test was determined. The sensitivity of the ELISA relative to the IFA test was 82.64%, and the specificity of the ELISA relative to the IFA test was 56.25%. Agreement between the ELISA and the IFA test was highly significant (Kappa = 0.74, Z = 5.78). We concluded that the ELISA is as good as the IFA test for detecting CAA antibody in sera from chickens.     

鸡白痢、鸡伤寒沙门菌分子分型方法的验证

为验证鸡白痢、鸡伤寒沙门菌分子分型方法的准确性,本研究以3株鸡白痢、鸡伤寒沙门菌国际参考菌株和306株国内分离株为研究对象,采用5种已知的分子分型方法开展验证性实验.结果发现,基于特定基因可变区的2种分型方法具有良好的特异性,其检测结果与生化分型结果一致,而3种基于特定基因单核苷酸多态性的分型方法则出现假阳性或假阴性的...     

Detection of antibodies to Mycoplasma gallisepticum vaccine ts-11 by an autologous pMGA enzyme-linked immunosorbent assay

Mycoplasma gallisepticum is a poultry pathogen that causes respiratory disease and loss of egg production worldwide. A live attenuated vaccine, ts-11, has been used for control of M. gallisepticum in several countries. The rapid serum agglutination test is usually used as an indicator of flock response to vaccination; however, in some flocks, the detected response may be weak or absent. With the use of specific monoclonal antibodies against M. gallisepticum strain S6 pMGA in immunoaffinity purification, the major membrane antigen of ts-11 was purified. An indirect enzyme-linked immunosorbent assay (ELISA) was developed with the purified antigen, and its potential for detection of antibodies induced after ts-11 vaccination was compared with an indirect ELISA with M. gallisepticum strain S6 pMGA. In the presence of high levels of ts-11-induced antibodies, both antigens detected similar numbers of positive sera. However, when lower levels of antibodies were present, ts-11 pMGA showed a higher sensitivity than S6 pMGA. Further examination of ts-11 pMGA with Mycoplasma synoviae-infected chicken sera revealed that ts-11 pMGA is specific for M. gallisepticum antibodies. With a panel of sera from ts-11-vaccinated or non-ts-11-vaccinated field chickens, the ts-11 pMGA ELISA was found to be more sensitive than the commercial rapid serum agglutination test in detecting antibodies to ts-11 vaccine. The results from this study suggest that the major membrane antigen of M. gallisepticum may have slightly different antigenic profiles in different strains, thereby necessitating the use of autologous antigens in serodiagnostic assays to increase sensitivity of the tests for mycoplasma antibodies. Thus, the low level of antibody response after ts-11 vaccination is, at least partially, due to the low ability of the current diagnostic antigens to bind ts-11 antibodies.     

重组N蛋白抗原检测PRRSV抗体ELISA的研究Ⅱ.试剂盒阴阳性临界值的测定及其与IDEXX公司试剂盒的比较

对临床上采集的899份猪血清样本。用以纯化的重组N蛋白为包被抗原建立的检测猪繁殖与呼吸综合征病毒（PRRSV）抗体的间接ELISA进行检测．运用统计学方法摸清了检测结果的分布规律。并扣国外IDEXX公司PRRSV抗体检测试剂盒同时对460份血清样本进行检测。结果表明。2种方法的符合率为91．73％。利用TG—ROC软件确定了自制的ELISA酶标板（NP—ELISA）的临界值。并标定试剂盒的特异性扣敏感性均为92．6％。ELISA的结果判定标准是：当以血清样本L为标准参考阳性血清时，样品与阳性血清的比值（S／P）小于或等于0．4为阴性；S／P在0．4与0．5之间为可疑；S／P大于或等于0．5为阳性。与IDEXX公司PRRSV抗体检测试剂盒对临床样品的检测结果进行比较后，初步判定所建立的ELISA之所以出现较多的假阴性。可能是目前临床上出现了PRRSV欧洲型所致。