鸡白痢与鸡伤寒沙门菌双重PCR方法的建立和初步应用

Detection of Salmonella Gallinarum and Pullorum strains by double PCR

分析不同血清型/生物型沙门菌全基因组序列筛选出鸡白痢和鸡伤寒沙门菌特异性基因组序列,设计两对引物,建立双重PCR方法鉴别检测鸡白痢和鸡伤寒沙门菌不同生物型,并进行初步的临床应用。双重PCR方法结果显示,鸡白痢沙门菌显示417 bp条带,鸡伤寒沙门菌显示417 bp和636 bp两个条带,而阴性对照未出现条带,与预期设计相符。双重PCR体系对56株不同血清型沙门菌鉴定结果与细菌学分离的血清型鉴定结果完全一致,说明本试验建立的双重PCR体系特异性良好,应用上述方法检测鸡场疑似20份临床样本,结果发现8株鸡白痢沙门菌阳性,1株鸡伤寒沙门菌阳性。上述结果表明,已建立双重PCR方法特异性检测鸡白痢和鸡伤寒沙门菌。本试验为鸡白痢和鸡伤寒不同生物型沙门菌的检测提供了一种简洁、敏感、特异的新方法。

Specific genes for Salmonella Gallinarum and Pullorum wero screened by analysis of different salmoneia serotypes/biovars wholo genome sequences.Two pairs of primer were designed and double-PCR was established te identify Salmonella Gallinarum and Pullorum,and then used for detecting cliniccl samples.The results showed that the Salmonella Pullorum had a 417 bp band,and salmonella Gallinauum exhibited both 417 bp and 636 bp.Double-PCR results showed the right Salmonella Gallinauum and Pullorum strains form 56 different Salmonella serotype strains.This method was applied te detect 20 clinical samples and 8 SalmonXm Pullorum trains and 1 Salmonella Gallinauum strain were found.The above results indicate that the method is a simple and specific way for Salmonella Gallinarum,and Pullorum detection.