CAJ | 학술논문

建立基于Taqman探针实时荧光PCR检测鸡伤寒沙门氏菌（Salmonella galli-narum）的方法．根据GenBank公布的鸡伤寒沙门氏菌基因序列，设计引物和Taqman探针，采用实时荧光PC R进行特异性、灵敏性及模拟样品的检测实验．结果表明：特异性引物和探针可从34株鸡伤寒沙门氏菌菌株、26株其他血清型沙门氏菌和7株非沙门氏菌菌株中检测出全部的34株鸡伤寒沙门氏菌．以鸡伤寒沙门氏菌梯度稀释菌液DNA为模板进行实时荧光PCR实验，菌株模板浓度与Ct值呈良好线性关系，线性系数（R2）为0.999，扩增效率为88．2％，最低检测浓度为5cfu/mL．对已接种鸡伤寒沙门氏菌的模拟样品同时进行实时荧光PCR检测和传统方法鉴定，两者结果一致．该方法特异性好、灵敏度高，是快速鉴定鸡伤寒沙门氏菌的有效方法．
건립기우Taqman탐침실시형광PCR검측계상한사문씨균（Salmonella galli-narum）적방법．근거GenBank공포적계상한사문씨균기인서렬，설계인물화Taqman탐침，채용실시형광PC R진행특이성、령민성급모의양품적검측실험．결과표명：특이성인물화탐침가종34주계상한사문씨균균주、26주기타혈청형사문씨균화7주비사문씨균균주중검측출전부적34주계상한사문씨균．이계상한사문씨균제도희석균액DNA위모판진행실시형광PCR실험，균주모판농도여Ct치정량호선성관계，선성계수（R2）위0.999，확증효솔위88．2％，최저검측농도위5cfu/mL．대이접충계상한사문씨균적모의양품동시진행실시형광PCR검측화전통방법감정，량자결과일치．해방법특이성호、령민도고，시쾌속감정계상한사문씨균적유효방법．
A method was developed for rapid identification of detecting Salmonella gallinarum by real-time fluorescent PCR .According to the gene of Genbank ,a set of primers and Taqman probe was de-signed to perform specific ,sensitive and simulation sample tests by real-time PCR .The results were confirmed that the specificity probe correctly distinguished 34 Salmonella gallinarum strains from 24 other Salmonella serotypes strains and 7 non-Salmonella strains .Gradient dilutions of Salmonella gallinarum were used as template to perform real-time PCR assay which presented linear relationship between the concentration of template and Ct value .Linear coefficient (R2 ) ,efficiency and detection limit were 0 .999 ,88 .2% and 5 cfu/mL correspondingly ,simulation samples inoculated with Salmo-nella gallinarum were detected by real-time PCR assay .The PCR method yielded a 100% correlation with results obtained by conventional culture method .The newly method showed that a high specific-ity ,sensibility and accuracy could be applied for rapid identification of Salmonella gallinarum .