CAJ | 학술논문

建立实时荧光PCR快速鉴定伤寒沙门氏菌（Salmonella Typhi）的方法。根据GenBank公布的伤寒沙门氏菌基因序列，设计引物和Taqman探针，采用实时荧光PCR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31株伤寒沙门氏菌菌株、27株其他血清型沙门氏菌和7株非沙门氏菌菌株中鉴定出全部的31株伤寒沙门氏菌。以伤寒沙门氏菌梯度稀释菌液DNA为模板进行实时荧光PCR扩增，菌株模板浓度与Ct值呈良好线性关系，线性系数为0.994，扩增效率为94.5%，最低检测浓度为4cfu/mL的添加浓度。实时荧光PCR检测与传统方法相比较，两者结果一致。该方法特异性好、灵敏度高，可以快速鉴定伤寒沙门氏菌。
건립실시형광PCR쾌속감정상한사문씨균（Salmonella Typhi）적방법。근거GenBank공포적상한사문씨균기인서렬，설계인물화Taqman탐침，채용실시형광PCR진행특이성、령민성급모의양품적검측실험。결과표명，특이성인물화탐침가종31주상한사문씨균균주、27주기타혈청형사문씨균화7주비사문씨균균주중감정출전부적31주상한사문씨균。이상한사문씨균제도희석균액DNA위모판진행실시형광PCR확증，균주모판농도여Ct치정량호선성관계，선성계수위0.994，확증효솔위94.5%，최저검측농도위4cfu/mL적첨가농도。실시형광PCR검측여전통방법상비교，량자결과일치。해방법특이성호、령민도고，가이쾌속감정상한사문씨균。
A method was developed for Rapid Identification of Salmonella Typhi with real-time fluorescent PCR. According to the gene of Genbank, a set of primers and Taqman probe was designed to perform specific, sensitive and simulation sample tests with real-time PCR. The results showedthe specificity probe correctly distinguished 31 Salmonella Typhi strains from 27 other Salmonella serotypes strains and 7 non-Salmonella strains. Gradient dilutions of Salmonella Typhi 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 0.994, 94.5%and 4cfu/ml correspondingly. Simulation samples inoculated with Salmonella Typhi were detected with real-time PCR assay. The PCR method yielded a 100%correlation with results obtained by conventional culture method. The new method that showed a high specificity, sensibility and accuracy could be applied for the rapid identification of Salmonella Typhi.