CAJ | 학술논문

根据NCBI沙门氏菌属invA基因保守序列设计引物，建立NASBA检测方法，进行特异性试验和灵敏度试验，并采用人工添加菌株污染样品进行方法验证。结果表明，引物SM-F/SM-R特异性强，能够从污染样品中扩增出277 bp的特异性片段，试验中未出现假阳性和假阴性结果；灵敏度高，对目标菌检出限达到5.5 pg/μL总RNA；检测周期短，NASBA反应在42℃90 min即可进行有效扩增，24 h内可完成样品中目标菌的筛选。NASBA技术适用于基层检验机构和现场快速检测食源性致病菌。
근거NCBI사문씨균속invA기인보수서렬설계인물，건립NASBA검측방법，진행특이성시험화령민도시험，병채용인공첨가균주오염양품진행방법험증。결과표명，인물SM-F/SM-R특이성강，능구종오염양품중확증출277 bp적특이성편단，시험중미출현가양성화가음성결과；령민도고，대목표균검출한체도5.5 pg/μL총RNA；검측주기단，NASBA반응재42℃90 min즉가진행유효확증，24 h내가완성양품중목표균적사선。NASBA기술괄용우기층검험궤구화현장쾌속검측식원성치병균。
A nucleic acid sequence-based amplification (NASBA)for the detection of salmonella was established using primers specific to invA gene. Specificity and sensitivity test was carried out and artificially contaminated samples were used to verify the method. The assay could specifically amplify a 277 bp fragment from artificially contaminated samples by primer SM-F/SM-R , while no false positive or false negative results were happened. The detecting limit of NASBA was 5.5 pg/μL nucleic acid. The assay could amplify effetely on condition of 42℃90 min.The detection of Salmonella could be completed in 24 hours. The NASBA assay was proved to be compatible for the base course and also for the field inspection rapidly.