Research and Application on Detection of Salmonella sp. by FQ-PCR

Abstract:

The FQ-PCR has been introduced for the Salmonella spp. detection these years. To develop a FQ-PCR assay to rapid,sensitive, specific and accurate for quantitative detect Salmonella spp. A pair of specific primers and probe were designed from the fimY gene of Salmonella spp. in this research. Through optimizing the Taq enzyme、Mg²⁺ 、primer concentration and probe concentration system and condition of FQ-PCR, the detection sensitivity was improved dramatically. The results of optimum experiment as following:Taq enzyme:2.5U;Mg²⁺concentration:3.75×10^-3mol/L;primer concentration 0.65×10^-6mol/L;probe concentration 0.30×10^-6mol/L;Cycle condition: step1:95℃,2min,step2:95℃ 5s,60℃ 40s,40cycles. Not only is this detection technology more specific, sensitive and efficient, but also the processing is wide application, rapid and simple. This FQ-PCR detection technology, therefore,will be used to detect Salmonella spp. for those areas food sanitation supervision, commodity inspection and detection,clinical diagnosis, etc.