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本研究旨在对吉林省延吉市三道湾镇两位农户饲养的病死黄牛进行快速诊断。本试验采集肝脏、脾脏、心脏、肌肉等组织,通过流行病学调查、临床剖检、显微镜观察、细菌培养观察、生化试验、分子生物学试验及动物试验进行气肿疽病诊断。病死黄牛的肌肉部位触压有捻发音,切面有大量血液和气泡流出,镜检可见两端钝圆、有芽孢的大杆菌,在肝片肉汤培养基中形成上清清朗、底部有松散的白色沉淀,生化试验均呈现气肿疽厌气菌所特有的产酸产气反应,PCR扩增出大小为501 bp的特异性条带。结果表明延边地区两病死黄牛感染了气肿疽病,证明了该地区气肿疽梭菌的存在。本试验首次分离鉴定出气肿疽梭菌延边株,为该地区气肿疽病的诊断和防制提供了重要的参考依据,同时为进一步研究该病的药物防制和免疫防制奠定了重要基础。 相似文献
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DUAN Xue-yan JIN Dong-chun REN Chun-yu CHE Da YUN Jin-yan QI Qiang JIN Xin 《中国畜牧兽医》2015,42(10):2800-2805
The study was aimed to rapidly diagnose two ill and dead cattle in three bay town of Yanji city in Jilin province.We collected liver,spleen,heart,muscle and other tissues to diagnose Clostridium chauvoei infection by epidemiological investigation,clinical autopsy,microscopic obsevation,bacterial culture observation,biochemical experiment,molecular biology diagnosis and animal experiment.The muscles of the ill and dead cattle contacted with crepitus,and the section had a lot of blood and bubble flowing out,microscopic obsevation found bacillus with obtuse at both ends and spores,the bottom of the tube had loose and white precipitate in the broth,biochemical experiments showed that the isolate had acid-production and gas-production reactions which was unique to anaerobic bacteria causing Clostridium chauvoei infection,and 501 bp fragment was amplified by PCR.The results showed that two ill and dead cattle were casued by Clostridium chauvoei infection in Yanbian area,and proved the existence of Clostridium chauvoei.The test separated Clostridium chauvoei Yanbian strain for the frist time,which provided important reference basis for the prevention and treatment of Clostridium chauvoei infection,and laid the important foundation for the further study of drug and immune preventions of this disease. 相似文献
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二重PCR方法鉴别气肿疽梭菌和腐败梭菌 总被引：1，自引：1，他引：0
用包含16S～23S rDNA间隔区和23S rDNA的部分序列作为气肿疽梭菌特异性标志,以α毒素部分序列作为腐败梭菌特异性标志,建立了鉴别气肿疽梭菌和腐败梭菌的二重PCR方法。结果显示：气肿疽梭菌C54-1株扩增出大小为509 bp的条带,腐败梭菌C55-1株、C55-16株均扩增出大小为148 bp的条带,均与预期吻合;而产气荚膜梭菌C57-1株、C59-37株,肉毒梭菌C62-4株,诺维梭菌C61-4株均未扩增出任何条带。扩增产物的测序结果进一步证实了本方法的特异性。菌株的生物学特性试验结果也符合相应气肿疽梭菌和腐败梭菌的特点。本研究所建立的二重PCR方法可用于气肿疽梭菌和腐败梭菌的快速鉴定。 相似文献
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Summary The Enzyme Linked Immunosorbent Assay (ELISA) provides a very efficient technique for detecting antibodies against Dictyocaulus vivparus in calves. Although low level cross reactions were found in animals with gastrointestinal nematodes, the specificity and sensitivity of the technique are sufficient for herd diagnosis of lungworm infections and for survey work. This conclusion is reached on the basis of artificially and naturally infected calves. ELISA titres correlate well with Indirect Haemagglutination titres, parasitological findings, and clinical observations. 相似文献
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[目的]构建联合表达气肿疽梭菌的鞭毛蛋白与分泌蛋白神经氨酸酶的方法,用以制备气肿疽生物工程亚单位疫苗.[方法]根据已公布的气肿疽梭菌FliA（C）和NanA序列设计引物,联合使用SOE-PCR与TD-PCR技术,建立融合基因FliA（C）-NanA的扩增方法.[结果]试验成功扩增出了融合基因FliA（C）-NanA.[结论]SOE-PCR方法无需在引物设计中加入酶切位点进行连接,降低了试验成本,且2个基因间未加入其他核酸序列,避免了表达出影响免疫效果的蛋白;TD-PCR的应用节省了试验时间,避免了筛选退火温度的繁琐步骤,解决了基因丢失的问题. 相似文献
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[目的]了解气肿疽梭菌的抗原特性。[方法]对气肿疽梭菌标准菌株(C54-1)的菌体抗原进行提纯过柱,通过制备小鼠免疫血清及免疫印迹技术进行了反应原性试验。[结果]气肿疽梭菌的菌体抗原有8条蛋白带,电泳图谱上分别出现了与之相对应的条带,分子质量分别为93、72、56、46、38、26、15、11 kD,其中第2(72 kD)、5(38 kD)、6(26 kD)峰的抗原表现出明显的特异性。用酶联免疫印迹技术分析气肿疽梭菌分泌蛋白的抗原组分,气肿疽梭菌感染小鼠的血清能与抗原组分蛋白发生反应,其中72、38、26 kD蛋白带显色最深。[结论]为气肿疽梭菌的深入研究提供了理论依据。 相似文献
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