In this study, a real-time loop-mediated isothermal amplification (Rti-LAMP) combined with immunomagnetic beads separation (IMS) assay and SDS rinse was developed for the rapid and sensitive detection of Salmonella in ready-to-eat (RTE) meat without prior enrichment. Polyclonal antibody and specific immunomagnetic beads (IMBs) for Salmonella flagellum were prepared. The capture efficiency was more than 95% by adding 50 μL IMBs into 1 mL of Salmonella suspension for absorption at 37 °C for 20 min. Then, Gly-HCl was the key to the effective eluting and separating of bacteria from IMB-bacterial complexes. The eluted bacteria were resuspended with 1× TZ lysing solution and boiled at 99.5 °C for 10 min, as DNA template of Rti-LAMP assay. The development of IMS-Rti-LAMP assay could specifically detect Salmonella as low as 5 CFU/mL in 0.85% saline solution. The lowest level of Salmonella DNA consistently detected by the IMS-Rti-LAMP assay was that from 75 CFU per 25 g (3 CFU/g) of RTE duck neck meat. The whole assay could be completed in 3 h. In addition, the IMS-Rti-LAMP assay was used to detect 22 RTE duck meat samples from local market, using plate culture as control. The result showed that four samples were positive for Salmonella by IMS-Rti-LAMP assay; three of them were also positive by plate culture method. These results showed that the development of IMS-Rti-LAMP combined with SDS rinsing was a rapid and sensitive method for the detection of Salmonella in RTE duck meat without enrichment.

在这项研究中，实时定量环介导的等温扩增（Rti-LAMP）与免疫磁珠分离（IMS）分析和SDS漂洗相结合，可用于快速，灵敏地检测即食（RTE）肉中的沙门氏菌无需事先充实。制备了沙门氏菌的多克隆抗体和特异性免疫磁珠（IMB）。通过将50μLIMB加入1 mL沙门氏菌中，捕获效率超过95％悬浮液，在37°C下吸收20分钟。然后，Gly-HCl是有效洗脱和从IMB-细菌复合物中分离细菌的关键。将洗脱的细菌重悬于1x TZ裂解液中，并在99.5°C下煮沸10分钟，作为Rti-LAMP分析的DNA模板。IMS-Rti-LAMP测定法的发展可以特异性检测0.85％盐溶液中低至5 CFU / mL的沙门氏菌。IMS-Rti-LAMP分析一致地检测到的最低沙门氏菌DNA含量为每25克RTE鸭脖子肉中75克分子毒素（CFU）。整个测定可以在3小时内完成。此外，使用平板培养作为对照，使用IMS-Rti-LAMP分析法检测了来自本地市场的22个RTE鸭肉样品。结果显示，四个样本对IMS-Rti-LAMP检测沙门氏菌；通过平板培养法，其中三个也呈阳性。这些结果表明，IMS-Rti-LAMP结合SDS冲洗的开发是一种无需富集即可快速检测RTE鸭肉中沙门氏菌的方法。