鸡白痢沙门氏菌与其它致病性沙门氏菌的血清型特异性PCR鉴别检测方法的建立

摘要

To establish a protocol for rapid differential detection of Salmonella pullorum from other pathogenic Salmonella serotypes,a PCR method for specific detection of S.pullorum was developed with a pair of primers targeting the SEEP17495 gene of S.pullorum.Under the optimized reaction conditions,the detection results showed that this method was able to amplify the 356 bp specific DNA fragment from S.pullorum,but no any amplification was found for 29 other pathogenic Salmonella serotypes and 7 common non-Salmonella pathogens.In addition,the detection limit of the method was 103 cfu/mL from both pure bacteria culture and the chicken manure samples.Therefore,this detection method established in this study showed high specificity and sensitivity,which provided a rapid and effective method for the S.pullorum detection.%为建立鸡白痢沙门氏菌与其他常见致病性沙门氏菌的血清型特异性快速PCR检测方法,本研究通过对GenBank中鸡白痢沙门氏菌、鸡伤寒沙门氏菌和肠炎沙门氏菌全基因组进行生物信息学分析,确定SEEP17495基因为鸡白痢沙门氏菌的检测靶基因,设计特异性引物,建立了一种能够直接从粪便样品中检测鸡白痢沙门氏菌的PCR检测方法.结果显示:该方法对于两株鸡白痢沙门氏菌均可以特异性地扩增出356 bp的目的片段,但对于7株常见非沙门氏菌致病菌和29株其他常见致病性沙门氏菌血清型的扩增结果均为阴性.该方法无论检测鸡白痢沙门氏菌纯培养菌,还是检测模拟鸡粪样品中的鸡白痢沙门氏菌,检测的灵敏度均为103cfu/mL.本研究建立的PCR检测方法具有良好的特异性和灵敏性,并且能够快速检测出粪便样品中的鸡白痢沙门氏菌,为鸡白痢沙门氏菌的检测提供了一种快速、有效的方法.