Rapid Detection of Cronobacter muytjensii by Real-time PCR Based on cgcA Gene
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Abstract:
To achieve the rapid detection of the common contaminating bacteria C. muytjensii in milk powder, specific probes and primers were designed based on the cgcA gene. The reaction system and reaction conditions for the detection of C. muytjensii were developed using quantitative real-time polymerase chain reaction (PCR) assay. Additionally, the specificity, sensitivity, and reproducibility of this method were evaluated, and the method was applied for the detection of artificially contaminated samples. The results showed that the specificity of real-time PCR assay was 100%; the sensitivity results revealed a detection limit of 440 cfu/mL for C. muytjensii. The reproducibility test results showed that the intra-assay coefficient of variations (CVs) ranged from 0.48% to 0.69%, while inter-assay CVs ranged from 0.69% to 0.73%. For the artificially contaminated samples, a positive result was obtained after 6 h of enrichment. The established real-time PCR assay exhibited good specificity, high sensitivity, and high reproducibility, and has promising potential to be a novel method for the rapid detection of C. muytjensii in food as well as a good value in research and application.
