鸡白痢沙门氏菌PCR检测技术的建立与应用

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。

Construction and Application of PCR Detection Technology on Salmonella Pullorum

Based on the published sequences of the Salmonella pullorum fliC gene in GeneBank,a pair of primers was synthesized.A Salmonella strain and other three non-Salmonella strains were extracted DNA and detected by Polymerase Chain Reaction（PCR）.Through sensitivity testing,the results showed that the Salmonella specific PCR product occurred 600bp band,rather than others did not occur,which proved the primers were specific.PCR can detect more than 50pg DNA.Using the above techniques,10 feed samples and 12 suspected disease samples were determined by PCR,the results showed that the positive detection rate and sensitivity higher than conventional detection methods.Thus,PCR detection of Salmonella is a rapid,sensitive and highly specific diagnostic method.