期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 412, 期 1, 页码 93-101出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-019-02209-y
关键词
Vibrio parahaemolyticus; Polymerase spiral reaction; Isothermal nucleic acid testing; Rapid detection
资金
- National Natural Science Foundation of China [81502849, 81872668]
- Bethune Medical Scientific Research Fund Project of Jilin University [2018B20]
- Scientific and Technological Research Project of Jilin Province [20170204003SF, 20180101095JC]
- Health science and technology capacity improvement project of Jilin Province [2019Q011]
- Fundamental Research Funds for the Central Universities
The aim of this study was to develop an effective and specific visual method to rapidly detect and identify Vibrio parahaemolyticus (V. parahaemolyticus) based on the polymerase spiral reaction (PSR). The method utilized only two pairs of primers designed specifically to target the conserved tlh gene sequence of V. parahaemolyticus. Nucleic acid amplification can be achieved under isothermal conditions using DNA polymerase. The reaction could be accomplished in < 40 min with high specificity and sensitivity. The limits of detection of V. parahaemolyticus in purified genomic DNA and pure culture were 300 fg/mu L and 2.4 CFU/mL per reaction, respectively, which were 100-fold more sensitive than with conventional PCR. The model food samples showed consistent specificity and sensitivity to the pure bacterial culture. With these encouraging results, it is expected that the novel, effortless and reliable isothermal nucleic acid testing assay developed in this study has potential to be applied to screening for V. parahaemolyticus in seafood samples.
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