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1.
鸡白痢抗原生产菌种变异菌落特性的鉴定康凯中国兽药监察所北京１０００８１近几年，我们在制备鸡白痢抗原标准品和冻干鉴定抗原生产用菌种时发现，编号为Ｃ７９－１的标准型鸡白痢沙门氏菌菌株在普通琼脂平板上出现变异的大菌落，有时这种变异大菌落占菌落总数的２０％。... 相似文献
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从郑州某鸡场死亡雏鸡的肝脏、脾脏和肾脏中分离纯化细菌，通过致病性试验、生化试验，并结合临床症状、病理变化和培养特性鉴定为鸡白痢沙门氏菌。经传代分离株、培养细菌、收集细菌、灭活细菌、浓缩抗原、调整抗原浓度、无菌检查及染色制备鸡白痢沙门氏菌平板凝集抗原，建立了检测鸡白痢血清抗体的微量平板凝集法。该方法具有特异性强、操作简便快速的优点。 相似文献
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缺失rfaH基因的鸡白痢沙门氏菌株的构建及鉴定 总被引：1，自引：0，他引：1
为研制可以鉴别检测基因缺失型鸡白痢疫苗与自然感染菌,本研究通过基因同源重组技术,利用高效自杀性载体系统,敲除鸡白痢沙门氏菌(S.pullorum)CVCC 79201株的rfaH基因,同时未引入任何抗生素抗性基因等外源DNA序列,经筛选获得重组S.pullorum(rSp)株。检测结果显示缺失菌由光滑型转变为粗糙型。rSp在普通琼脂培养基上传15代后,仍然保持粗糙型表型。动物试验结果表明,rSp免疫伊莎褐蛋鸡后,其抗血清可通过平板凝集试验与CVCC 79201免疫的血清相区分。本研究为S.pullorum缺失疫苗的研制提供了技术平台。 相似文献
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从郑州某鸡场死亡雏鸡的肝脏、脾脏和肾脏中分离纯化细茵,通过致病性试验、生化试验,并结合临床症状、病理变化和培养特性鉴定为鸡白痢沙门氏菌.经传代分离株、培养细菌、收集细菌、灭活细菌、浓缩抗原、调整抗原浓度、无菌检查及染色制备鸡白痢沙门氏菌平板凝集抗原,建立了检测鸡白痢血清抗体的微量平板凝集法.该方法具有特异性强、操作简便快速的优点. 相似文献
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琼脂扩散法排除平板凝集法中的假阳性鸡白痢 总被引：1，自引：0，他引：1
<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 相似文献
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新疆地区鸡伤寒白痢沙门氏菌病的调查 总被引：5，自引：0，他引：5
对新疆地区鸡沙门氏菌病进行了病原学、血清学和流行病学调查，结果表明分离的246株细菌与鸡白痢鸡伤少门氏菌诊断血清呈阳性反应；对15株鸡沙门氏菌进行抗原结构分析，表明8株为标准型鸡白痢伤寒沙门氏菌，7株为中间型鸡沙门氏菌，对其中86株菌的生化鉴定表明符合鸡白痢鸡伤寒沙门氏菌的生化特性。该病在我区的发病死亡率为1％－25％，发病严重的鸡场为50％－75％。该病除了单一感染外，还与鸡大肠杆菌、葡萄球菌、鸡球虫病混合感染。 相似文献
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大肠杆菌菌体O抗原单因子定型血清的制备 总被引：1，自引：0，他引：1
为研制大肠杆菌菌体O抗原单因子定型血清,以大肠杆菌参考菌株CVCC1345(O2)、CVCC1350(O7)、CVCC1414(O74)为菌种制备抗原免疫家兔,获得了定型原血清,并通过比较不同免疫程序及剂量优化了定型原血清的制备工艺。进而按照优化后的定型原血清制备工艺,以大肠杆菌参考菌株CVCC1343(O1)、CVCC1345(O2)、CVCC1350(O7)、CVCC1405(O64)、CVCC1414(O74)、CVCC1439(O100)、CVCC1442(O103)、CVCC1455(O116)、CVCC1466(O128)、CVCC3801(O154)为菌种制备抗原免疫家兔,获得了10种O抗原型定型原血清。将定型原血清分别与180种微量凝集试验抗原逐一反应,明确各原血清与非特异性抗原的交叉凝集价,通过用吸收抗原对原血清进行吸收及稀释的方法消除非特异性凝集,最终制备成大肠杆菌菌体O抗原单因子定型血清。血清质量评价结果表明,新制备的单因子定型血清性状为无色、淡黄色或黄色澄明液体,个别有少量絮状物;无菌生长;特异性良好;凝集效价为1︰2。研究结果显示,制备的大肠杆菌O抗原单因子定型血清具有较好的特异性,可以用于对临床分离大肠杆菌进行O抗原血清型鉴定。 相似文献
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为建立活的非可培养状态(VBNC)研究模型,本研究利用液体LB和4℃联合条件对鸡白痢沙门氏菌CVCC578参考株的进行VBNC诱导,构建VBNC研究模型。同时依靠胎牛血清和程序性升温对处于该状态的菌体进行复苏,并对复苏前后的细菌进行了16SrRNA验证。结果表明:实验菌株经液体LB和4℃联合诱导后,可培养菌数在55d后降至零,总菌数在整个观察期内基本不变,而活菌数在150d后开始下降,180d后下降显著,表明实验菌株可在55d进入VBNC,而且维持时间至180d。当进入该状态后,菌体形态可由杆状变为球杆或球形,并且菌体排列可由单在变为聚集。经复苏和16SrRNA鉴定后,"变态"的细菌被证实为沙门氏菌,而非杂污染菌。该实验为规范VBNC沙门氏菌的鉴定程序以及制订相应的国家检测标准提供了实验依据。 相似文献
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1例鸡白痢沙门氏菌的分离鉴定与耐药性分析 总被引：1，自引：1，他引：0
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。 相似文献
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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。 相似文献
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An allele-specific PCR assay for the rapid and serotype-specific detection of Salmonella pullorum 总被引：2，自引：0，他引：2
Desai AR Shah DH Shringi S Lee MJ Li YH Cho MR Park JH Eo SK Lee JH Chae JS 《Avian diseases》2005,49(4):558-561
Salmonella serovar Pullorum is a causative agent of pullorum disease (PD) in poultry and is responsible for severe economic losses to the poultry industry in many parts of the world. A definitive detection of Pullorum requires culture followed by serotyping and biochemical identification, a process that is tedious and takes several weeks to accomplish. We have developed a rapid allele-specific polymerase chain reaction (PCR) method based on the nucleotide polymorphism in rfbS gene sequence for the serotype-specific detection of Pullorum and its differentiation from the closely related Gallinarum. The specificity of this PCR assay was tested using DNA samples from Pullorum (n = 13), Salmonella serotypes other than Pullorum (n = 19), and closely related non-Salmonella organisms (n = 5). The PCR assay was highly serotype-specific as the PCR amplicon of 147 base pairs was observed only in the case of Pullorum, while all the other DNA samples tested PCR negative. A definitive identification of Pullorum cultures was possible in less than 3 hr. As little as 100 pg of SP DNA was detected. This allele-specific PCR method is highly specific as well as sensitive and may be an effective molecular tool in the rapid and serotype-specific detection of Pullorum and differentiation from other Salmonella species. 相似文献
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The structure and serological specificities of the lipopolysaccharides (LPSs) from Salmonella enterica serovar Gallinarum biovar Pullorum were studied to provide an improved basis for the distinction between antigenic types and the development of improved diagnostic tests. The structure of the LPS O-polysaccharide (O-PS) from S. Pullorum standard, intermediate and variant antigenic type strains was determined by mass spectrometry, nuclear magnetic resonance spectroscopy and chemical analysis. The LPS of the three strains shared a common structural repeating oligosaccharide unit containing d-mannose, l-rhamnose, d-galactose and d-tyvelose (1:1:1:1). The O-PS of the variant type LPS contained an additional d-glucose residue linked to the O-4 position of the d-galactose residue. The O-PS of the intermediate type LPS was partially the same as that of the variant LPS, however, the molar ratio of the d-glucose component was lower with respect to the other glycose components. Serological specificities of the three antigenic type LPSs were examined with anti-S. Pullorum LPS monoclonal antibodies (Mabs). On immunoblots, Mabs to the standard type O-PS reacted with high molecular mass (HMM) and low molecular mass (LMM) LPS from the standard strain, and with LMM but not HMM LPS from the variant strain. Monoclonal antibodies to the variant type O-PS reacted with HMM but not LMM LPS from the variant strain, and did not react with HMM or LMM LPS from the standard strain. On ELISA, the standard, intermediate and variant antigenic type strains were differentiated by the relative reactivity with the anti-LPS O-PS Mabs. Several of the anti-LPS O-PS Mabs were specific for S. Pullorum and other serogroup D1 Salmonella, and are potentially useful for the development of improved diagnostic tests for these organisms. 相似文献
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鸡白痢沙门氏菌PCR检测技术的建立与应用 总被引：1，自引：0，他引：1
为鸡沙门氏菌的临床检测提供了一种更快速、敏感、特异的诊断,参照GeneBank中已公布的的鸡白痢沙门氏菌fliC基因序列,合成出了一对引物,使用PCR法对1株沙门氏标准菌株和其他3株非沙门氏菌标准菌株进行DNA抽提扩增并检测其敏感度。采用上述技术,对12份可疑病料及10份饲料进行PCR检测,同时与传统检测方法进行比较。结果表明1株沙门氏菌PCR产物出现600 bp的特异性DNA扩增带,而非沙门氏菌均未出现扩增条带,证明所设计引物具有沙门氏菌特异性;通过敏感度检测,此PCR体系能检出50 pg以上的细菌DNA,敏感性较高。运用PCR法阳性检出率及敏感性均高于常规检测方法。由此可见,沙门氏菌PCR检测是一种快速、敏感和高度特异的诊断方法。 相似文献
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Serological response of chickens to infection with Salmonella gallinarum-S. pullorum detected by enzyme-linked immunosorbent assay. 总被引：1，自引：0，他引：1
The serological response to Salmonella pullorum and S. gallinarum infection in chickens was studied with an indirect enzyme-linked immunosorbent assay (ELISA). In broiler chickens, a more virulent strain of S. pullorum produced a significantly lower serum IgG titer than did a less virulent strain. In laying hens, the serum and egg-yolk IgG titers were very similar. In chickens infected with S. gallinarum, high IgG titers persisted for 30 weeks. In chickens reinfected with this strain, each reinfection was followed by transitory increases in IgG lasting no longer than 2 weeks. Serum samples from Brazil taken from a laying flock with evidence of fowl typhoid showed much higher antibody levels than did those from three uninfected flocks. Using lipopolysaccharide as the detecting antigen, infections caused by these salmonellae could be differentiated from those caused by other groups. Incorporation of the appropriate flagella antigen in the ELISA allowed differentiation between infections caused by S. pullorum and S. enteritidis. 相似文献
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Allgayer MC Lima-Rosa CA Weimer TA Rodenbusch CR Pereira RA Streck AF Oliveira SD Canal CW 《The Veterinary record》2008,162(25):816-819
Cloacal swabs were collected from 280 captive psittacine birds belonging to 13 species. Samples of dna were tested by PCR using a pair of primers that amplify a 284 base pair fragment of the Salmonella genus invA gene, and the PCR-positive samples were tested by standard microbiological techniques. Thirteen per cent of the samples were positive by PCR, but negative by microbiological techniques. The infection rates were significantly different among the 13 species, the most commonly infected being Amazona amazonica (28 per cent) and Amazona pretrei (20 per cent). Specific tests for Salmonella Typhimurium Salmonella Enteritidis, Salmonella Pullorum and Salmonella Gallinarum did not produce positive results. 相似文献
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LIU Zhi-ke YANG Ning-ning XU Jin-feng XU Ming-guo JING Ming-long WU Wen-xing CAO Xu-dong REN Yan SHI Feng CHEN Chuang-fu 《中国畜牧兽医》2017,44(9):2792-2800
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. 相似文献
20.
Salmonella pullorum is the cause of pullorum disease, which is characterized by white diarrhea and a high mortality rate in poultry. During the 1990s, the serologic "pullorum" test has occasionally failed to detect infected birds during the early stage of disease. To determine if any recent genetic changes have taken place in S. pullorum to account for poor seroconversion sometimes observed in infected flocks, S. pullorum from 1990s outbreaks and strains isolated prior to the 1980s were typed by random amplified polymorphic DNA (RAPD). Of 40 S. pullorum isolates typed by this method, eight distinct DNA patterns were identified with one of three RAPD polymerase chain reaction primers. Sixty-two percent of S. pullorum isolates shared the same RAPD DNA pattern, and a major proportion of these strains were from recent flock infections. The RAPD patterns for S. pullorum were clearly distinct from the avian Salmonella group B isolates included in this analysis. The distribution of Salmonella virulence genes among avian Salmonella isolates was also examined. Eighty-five percent of the S. pullorum isolates had both the virulence plasmid gene, spvB, and the invasion gene, invA, with the same percentage positive for the Salmonella enteriditis fimbrial gene, sef. However, significant variability was observed among S. pullorum in their ability to invade avian epithelial cells, despite the presence of the Salmonella invasion gene in these isolates. 相似文献