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Establishment of Loop-mediated Isothermal Amplification Assay for Detection of Salmonella

LI Jia-tong1, WANG Nan2, JIN Zhen-ai3, XU Yao4, SHAO Hong-ze4, JIN Xin1   

  1. (1.College of Agriculture, Yanbian University, Yanbian 133000, China;2.Jilin Province Center for Animal Disease Prevention and Control, Changchun 130062, China;3.Animal Health Supervision Institute of Hunchun City, Jilin Province, Hunchun 133300, China;4.Jilin Institute of Animal Husbandry and Veterinary Science Research, Changchun 130062, China)
  • Received:2013-08-16 Online:2014-03-20 Published:2014-05-15

Abstract: The assay was aimed to establish a rapid, sensitive and specific method of loop-mediated isothermal amplification (LAMP) for the detection of Salmonella. According to the highly conserved STN gene of Salmonella reported in GenBank, we designed a set of primers, and then followed by a series of LAMP optimization of reaction conditions, specificity test, sensitivity test, stability test and enzyme test. The sensitivity of LAMP and conventional PCR were compared.The results showed that the optimal reaction temperature of LAMP was 65 ℃, when only amplification of Salmonella, the minimum detectable concentration was 101 CFU/mL, which was more than conventional PCR detection by one order of magnitude. The results showed that LAMP method had the qualities of specificity, high sensitivity, short time-consuming and convenience for the detection of Salmonella, which was expected to develop into an effective method

Key words: loop-mediated isothermal amplification; Salmonella; detection; STN