关键词: 沙门氏菌, 鸡肉, 实时荧光定量环介导等温扩增, 检测

Abstract: In this study, a real-time loop amplified DNA assay system (Rti-LAMP) was developed for the rapid detection of Salmonella enterica on the chicken. It showed that the lowest level of 6 CFU/reaction for pure Salmonella culture could be detected and finished in 45 minutes by the Rti-LAMP targeting the invA gene using Midori Green as nucleic acid dye. Furthermore, seeding 25 g portions of chicken with various numbers of Salmonella CFU, followed enrichment culture at 37? C for 4 h, then filtration through Whirl-Pak bag and pelleting the bacterial cells by centrifugation at 13,000 g. The resulting pellets were suspended in saline and processed for cell lysis and DNA purification. 2 μL purified DNA samples was incorporated into 25 μL Rti-LAMP reactions at 65 ℃ for 60 min. The lowest level 450 CFU/g of Salmonella consistently detected by the Rti-LAMP assay. The entire assay could be completed in 7 h. 21 chicken samples purchased from market were detected by the Rti-LAMP assay as the control of Salmonella culture. The same one sample was Salmonella positive by the both of assays. It suggested that the sensitivity of the Rti-LAMP assay was as well as that of Salmonella culture on detection of chicken sample, but the former need only 7 h and was much more rapid than that of the latter.

Key words: Salmonella enterica ser. Enteritidis, chicken, real-time loop amplified, detection