CAJ | 학술논문

为了建立一种快速区分样品中沙门氏菌的死细菌与活细菌的检测方法，将叠氮溴化乙锭（EMA）与PCR技术相结合，以沙门氏菌的invA为靶基因，以沙门氏菌的纯培养细胞做模板进行扩增，并进行灵敏度、特异性、曝光时间及EMA浓度试验。结果显示，灵敏度为14 CFU/mL，最佳曝光时间为2 min，当EMA的浓度小于18μg/mL时，EMA对活菌靶基因的扩增没有明显的抑制，而终浓度为1μg/mL的EMA，能有效抑制1×108 CFU/mL沙门氏菌死菌的扩增。 EMA-PCR能有效降低沙门氏菌检测过程中的假阳性。
위료건립일충쾌속구분양품중사문씨균적사세균여활세균적검측방법，장첩담추화을정（EMA）여PCR기술상결합，이사문씨균적invA위파기인，이사문씨균적순배양세포주모판진행확증，병진행령민도、특이성、폭광시간급EMA농도시험。결과현시，령민도위14 CFU/mL，최가폭광시간위2 min，당EMA적농도소우18μg/mL시，EMA대활균파기인적확증몰유명현적억제，이종농도위1μg/mL적EMA，능유효억제1×108 CFU/mL사문씨균사균적확증。 EMA-PCR능유효강저사문씨균검측과정중적가양성。
In order to establish a fast detection method for distinguishing dead bacteria and live bacteria of Salmonella from samples, EMA and PCR were combined, meanwhile invA gene of Salmonella was taken as target gene, and then amplification was carried out by taking pure culture cells of Salmonella as template, furthermore sensitivity, specificity, exposure time and EMA concentration test were carried out. Results showed that:the sensitivity was 14 CFU/mL;the optimal exposure time was 2 min;when the concentration of EMA was less than 18 μg/mL, EMA had no obvious inhibition to the amplification of live bacteria′target gene, while EMA at final concentration of 1μg/mL could effectively inhibit the amplification of dead bacteria at 1×108 CFU/mL. So EMA-PCR could effectively decrease false positive rate in the detection of Salmonella.