Abstract
Vibrio parahaemolyticus (V. parahaemolyticus) is one of the most common and widely distributed food-borne bacteria. Thus, it is necessary to establish a rapid, specific, and sensitive detection approach for V. parahaemolyticus. The strategy of combining immunomagnetic separation method (IMS) and polymerase chain reaction (PCR) measure was established successfully for V. parahaemolyticus detection, because ordinary IMS always has restriction on employing conventional antibodies for their labor-intensive procedure, batch-to-batch variation in functionality, and limited shelf-life. Herein, we have generated a novel phage display-derived peptide with an amino acid sequence of MPRLPPA and use it as bio-recognition probe in IMS. The peptide was evaluated to have the advantages of high specificity, sensitivity, easy reproduction, and well-defined ability. When compared with the rabbit polyclone IgG antibodies (IgG)-IMS and chicken egg yolk antibodies (IgY)-IMS, peptide-IMS has absolute superiorities in terms of the dosages, detection time, and capture efficiency. It has the ability to take the place of traditional antibody and unquestionable value of reusing. Furthermore, the detection system of peptide-IMS-PCR possessed high specificity with a limit of detection (LOD) of 102 CFU/mL in pure culture and 103 CFU/mL in oyster. Thus, we successfully synthesize peptide against V. parahaemolyticus and applied it to establish a sensitive and specific detecting method for V. parahaemolyticus.
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Funding
This study was funded by the National Natural Science Foundation of China (Grant number 81502849, 81872668), the Scientific and Technological Research Project of Jilin Province (Grant numbers 20170204003SF and 20180101095JC), Health commission of Jilin Province (Grant number 2019Q011), the Bethune Medical Scientific Research Fund Project of Jilin University (Grant number 2018B20), and the Fundamental Research Funds for the Central Universities.
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Yue Zhai declares that he has no conflict of interest. Chao Zhao declares that he has no conflict of interest. Li Li declares that he has no conflict of interest. Kun Xu declares that he has no conflict of interest. Juan Wang declares that he has no conflict of interest. Xiuling Song declares that he has no conflict of interest. Hui Li declares that he has no conflict of interest.
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Zhai, Y., Zhao, C., Li, L. et al. Production of Phage Display-Derived Peptide and the Application for Detecting Vibrio parahaemolyticus by Combined PCR Technology. Food Anal. Methods 13, 1906–1917 (2020). https://doi.org/10.1007/s12161-020-01800-9
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DOI: https://doi.org/10.1007/s12161-020-01800-9