Scientia Agricultura Sinica ›› 2011, Vol. 44 ›› Issue (16): 3395-3402.doi: 10.3864/j.issn.0578-1752.2011.16.012

• STORAGE·FRESH-KEEPING·PROCESSING • Previous Articles     Next Articles

Development and Application of an IAC-PCR Kit for the Rapid Detection of Salmonella spp.

LI  Xiao-Ling, LIU  Bin, DAN  Xian-Long, WANG  Da-Peng, ZHOU  Min, SHI  Xian-Ming   

  1. 上海交通大学农业与生物学院/中美食品安全联合研究中心/陆伯勋食品安全研究中心
  • Received:2011-01-19 Revised:2011-04-21 Online:2011-08-15 Published:2011-05-04

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Abstract: 【Objective】The purpose of this study was to develop a rapid and accurate detection kit for Salmonella spp. using polymerase chain reaction (PCR) with an internal amplification control (IAC). 【Method】 The specific primers and IAC were designed according to the conserved genes invA and stn in Salmonella spp. The optimization of the components and the evaluation of the parameters for the kit were carried out in this study. 【Result】 The experiment indicated that the specific 362 bp DNA fragment was amplified against 147 reference strains of Salmonella spp., while 28 strains of non-Salmonella only showed the 520 bp amplified band of IAC. The detection limit of the kit for purified genomic DNA was 8.0 fg/PCR. It was confirmed in artificial contamination assay that Salmonella spp. could be detected by the kit after 8-10 h enrichment when 4-5 CFU germs were inoculated in 10 mL milk. Thirty food samples were detected by the IAC-PCR kit in this study, and the experiments demonstrated that IAC could successfully indicate false-negative results. Besides, the kit worked well after being frozen-thawed for 60 times or stored at -20℃ for one year.【Conclusion】The IAC-PCR detection kit developed in this study has good performances in specificity, sensitivity, stability and accuracy and is suitable for the rapid and accurate detection of Salmonella spp.

Key words: Salmonella spp., PCR, IAC, detection kit

CLC Number: 

  • T201

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