改良环介导等温扩增技术快速检测婴儿配方奶粉中的阪崎肠杆菌

Loop-mediated isothermal amplification assay for rapid detection of Enterobacter sakazakii in powdered infant formula

摘要:

摘要：【目的】采用改良环介导等温扩增（LAMP）技术，快速检测婴儿配方奶粉中的阪崎肠杆菌。【方法】以阪崎肠杆菌（ATCC29544）的16S-23S rRNA间区序列作为靶序列，设计内、外引物和环引物，通过肉眼观察白色沉淀，判断检测结果。【结果】LAMP检测阪崎肠杆菌的灵敏度为0.101 CFU/mL，人工污染阪崎肠杆菌的婴儿配方奶粉的检出限为1.1 CFU/g。采用试剂盒提取DNA，从样品处理到报告结果，耗时1 h。而对照，PCR检测阪崎肠杆菌的灵敏度为101 CFU/mL，人工污染阪崎肠杆菌的婴儿配方奶粉的检出限为1100 CFU/g。采用同样方法提取DNA，从样品处理到报告结果，耗时3 h。【结论】因此，LAMP检测婴儿配方奶粉中的阪崎肠杆菌灵敏度高，耗时短，方法简便。

Abstract:

Abstract: [Objective] A loop-mediated isothermal amplification (LAMP) technology with two loop primers was developed for rapidly detecting Enterobacter sakazakii in powdered infant formula. [Methods] Sequences of 16S-23S rRNA of Enterobacter sakazakii (ATCC29544) were used as target sequences, to design outer primers, inner primers and loop primers. We judged the results of detection, through visible to the naked eye of the white precipitate. [Results]The sensitivity of the LAMP assay was 0.101 CFU/mL; the detection limit of artificial contamination was 1.1 CFU/g. The detection could be finished about an hour from dealing with the sample to report the results with DNA kit. Compared with LAMP, the sensitivity of the PCR assay was 101 CFU/mL and the detection limit of artificial contamination was 1100 CFU/g in three hours. To apply the same method of extracting DNA, from dealing with the sample to report the results, it took about three hours. [Conclusion] Therefore, this LAMP-based assay is sensitive and fast. These results indicate that LAMP can provide a rapid yet simple test for the detection of Enterobacter sakazakii in powdered infant formula.