Production of Phage Display-Derived Peptide and the Application for Detecting Vibrio parahaemolyticus by Combined PCR Technology

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.