Production of Phage Display-Derived Peptide and the Application for Detecting Vibrio parahaemolyticus by Combined PCR Te
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Production of Phage Display-Derived Peptide and the Application for Detecting Vibrio parahaemolyticus by Combined PCR Technology Yue Zhai 1 & Chao Zhao 1 & Li Li 2 & Kun Xu 1 & Juan Wang 1 & Xiuling Song 1
&
Hui Li 1
Received: 30 January 2020 / Accepted: 14 June 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
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. Keywords Vibrio parahaemolyticus . Immunomagnetic separation method . Phage display-derived peptide . PCR
Introduction Vibrio parahaemolyticus, a Gram-negative halophilic microorganism, has been identified as a major food-borne pathogen of various seafoods throughout the world (Teng et al. 2017; Sakata et al. 2015). Human infections caused by V. parahaemolyticus often result from raw, under-cooked, or improperly processed molluscan shellfish, especially oysters (Lozano-Leon et al. 2003). When people ingest food Yue Zhai and Chao Zhao contributed equally to this work. * Xiuling Song [email protected] * Hui Li [email protected] 1
Department of Hygienic Inspection, School of Public Health, Jilin University, Changchun 130021, Jilin, People’s Republic of China
2
Department of Physiology, School of Basic Medical Sciences, Shenzhen University Health Sciences Center, Shenzhen 518060, People’s Republic of China
contaminated with V. parahaemolyticus, it might lead to acute gastroenteritis with nausea, headache, fever, or other serious complications, even including septicaemia (Raghunath et al. 2008; Zhang et al. 2015). Given the harmful effects and prevalence of V. parahaemolyticus, establishing a
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