Development of an Internal Amplification Control in the PCR Detection for Salmonella

Abstract

Abstract: By constructing an internal amplification control（IAC）, this study developed a PCR system for the detection of Salmonella, which could effectively indicate false-negative results. The specific primers were designed according to the conserved gene invA in Salmonella spp.. An IAC was constructed by the compound primer technology, and finally the PCR detection system was developed. The experiment indicated that the specific 385 bp DNA fragment was amplified against 33 reference strains of Salmonella spp., while 6 strains of non-Salmonella only showed the 484 bp amplified band of IAC. The detection limit of this PCR system forpurified genomic DNA was 6.35 fg/μL. The artificial contamination assays showed that Salmonella could be detected after eight hours enrichment when the original bacterial concentration was 3.2 CFU/25 mL. A large number of food samples were also tested, and the results demonstrated that the detection system could effectively avoid the false-negatives and improve the detection accuracy.

Key words: Salmonella, PCR detection, Internal Amplification, Control false-negative

Yang Jin, Zeng Qingmei, Zhang Di, Liu Kun, Wang Lin, Zhang Yajun. Development of an Internal Amplification Control in the PCR Detection for Salmonella[J]. Biotechnology Bulletin, 2014, 0(7): 54-59.

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