Detection of Weissella viridescens by Quantitative Real-Time Polymerase Chain Reaction Assay
ZHANG Yige1, JIANG Xiaobing1,*, YU Tao2, SUN Liying3
1.College of Life Sciences, Henan Normal University, Xinxiang 453007, China; 2.College of Life Sciences and Technology, Xinxiang University, Xinxiang 453003, China; 3.Foshan Haitian Flavouring and Food Co. Ltd., Foshan 528000, China
Abstract：A SYBR Green Ⅰ-based real-time polymerase chain reaction (PCR) assay for the detection of Weissella viridescens was established using rpoA as target gene in this study. Specific primers were designed targeting rpoA. The specificity, sensitivity, and repeatability of the real-time PCR method were evaluated and compared with those of conventional PCR. The real-time PCR method was found to be specific for W. viridescens. The limit of detection of this method was 2.667 × 10-3 pg/μL for genomic DNA from W. viridescens, and 30 CFU/mL and 0.8 CFU/g for pure bacterial culture and artificially inoculated beef, respectively. The sensitivity of real-time PCR was 1 000 times as high as that of conventional PCR. The results of independent repeated test on samples with different bacterial concentrations showed a standard deviation of less than 1 and coefficients of variation (CV) in the range of 0.02%–1.28% for cycle threshold (Ct). The real-time PCR method developed in this study can be considered to be a fast tool for detection of W. viridescens due to its good specificity, sensitivity, reproducibility, and accuracy.
2018, Vol. 32 
