Title: Establishment and Primary Application of Fluorescence Quantitive PCR for Detection of Vibrio parahaemolyticus
Abstract: Objective: The study developed a SYBR Green I real time quantitative PCR for Vibrio parahaemolyticus (VP). Method: According to genome sequences within toxR of VP published in GenBank, a pair of primers was designed by primer6.0. The reaction conditions, sensitivity and specificity of the method were optimized and evaluated. The method was also applied to clinical samples. Result: It was shown that the optimal primer concentration was 0.2 µmol/L. The results also demonstrated that standard curve established was shown a fine linear relationship between threshold cycle and template concentration. The correlation coefficient R 2 was 0.996, the efficiency of amplification in E was close to 100%, the dissolution curves were specific and the minimum detectable amount was 2.14 copies. This method has better specificity and reproducibility. The coincidence rate was 100% when compared the results to routine separation and assay method. Conclusion: A SYBR Green 1 fluorescent quantitative PCR for detecting toxR gene of VP was devel- noped which could be very useful for identification and inspection of VP from fishery product in import and export.
Publication Year: 2013
Publication Date: 2013-01-01
Language: en
Type: article
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