环介导恒温扩增(LAMP)—检测沙门氏菌

Detection of Salmonella.Spp by Loop-Mediated Isothermal Amplification(LAMP) Method

建立一种能够快速准确地检测沙门氏菌的LAMP方法。根据沙门氏菌invA基因设计了引物,然后进行LAMP反应条件的优化、特异性试验,通过LAMP与PCR灵敏度的试验与实际样品进行检出率的比较。LAMP方法特异性好,最佳反应温度为63℃,只对沙门氏菌进行扩增;沙门氏菌的检测灵敏度为7～8cfu/mL。LAMP方法检测沙门氏菌特异性强、灵敏度高、时间短且操作简便,有望成为快速检测沙门氏菌的新方法。

A rapid LAMP method was developed for detecting pathogen of Salmonella Spp. primers were designed according to invA gene of Salmonella Spp, optimization and specificity experiment of LAMP conditions was performed, and detection rate of LAMP assay and PCR method for actual samples was compared. The LAMP assay had high specificity, the optimized temperature was 63 ~C, and it only amplified Salmonella Spp; the sensitivity of LAMP assay for Salmonella Spp was 7-8 cfu /mL. The results suggested that detection of Salmonella Spp by LAMP was effective with high specificity and sensitivity, and was expected to become a valuable tool for rapid detection and identification of Salmonella Spp.