多重PCR检测病死鸡中沙门氏菌方法的研究
Study on the detection of ？？Salmonella？？ from dead chicken by multiplex PCR

为了建立能够同时检测病死鸡样品中的沙门氏菌、鸡白痢沙门氏菌和肠炎沙门氏菌的多重PCR方法. 采用煮沸法提取DNA做为模板, 选择分别针对沙门氏菌属、鸡白痢沙门氏菌和肠炎沙门氏菌的特异性基因(？？invA、fliC、sdfⅠ？？)设计引物, 优化反应体系和反应参数, 建立多重PCR检测方法. 应用该方法对国内17家鸡场369份病死鸡样品进行沙门氏菌的检测, 检测结果与传统细菌培养法的结果进行比较.结果表明, 优化过的多重PCR可同时对沙门氏菌种、属进行鉴定, 灵敏度为4.6×10？？？？2？？ CFU/mL；多重PCR方法共检测出沙门氏菌34株, 其中鸡白痢沙门氏菌21株、肠炎沙门氏菌5株, 与传统细菌培养法检测结果的符合率分别为91.2%、90.5%和80.0%.
To establish the multiplex PCR which could detect？？ Salmonella？？, ？？Salmonella pullorum？？ and ？？Salmonella enteritidis？？, simultaneously.The genomic DNA of bacteria was extracted by the boiling method, and the specific genes (？？invA, fliC, sdfⅠ？？) of ？？Salmonella？？, ？？Salmonella pullorum？？ and ？？Salmonella enteritidis？？ were selected to disign primers for establishing the multiplex PCR method , and then the specificity and sensitivity of multiplex PCR were tested; At the same time, 369 samples from 17 farms were detected by multiplex PCR, the results of the test were compared with the traditionaldetection methods. The results showed that the ？？Salmonella？？ species could be tested simultaneously by the optimized multiplex PCR, its sensitivity d was 4.6×10？？？？2？？CFU/mL; The number of ？？Salmonella？？, ？？Salmonellapullorum？？ and S？？almonella enteritidis？？ which detected by the multiplex PCR was 34, 21 and 5 , respectively, the coincidence rate with traditional bacteria culture method was 91.2%, 90.5% and 80.0% , respectively