APPLICATION OFMULTIPLEX POLYMERASE CHAIN REACTION TO THE DETECTION OF PATHOGENS IN FOOD LYNDA PERRY 1,5 , PRECIAUS HEARD 1 , MICHAEL KANE 2 , HANYOUP KIM 3 , SERGEI SAVIKHIN 3 , WILFREDO DOMÍNGUEZ 4 and BRUCE APPLEGATE 1 1 Department of Food Science 2 Department of Computer and Information Technology 3 Department of Physics Purdue University West Lafayette, IN 47907 4 Zamorano University Tegucigalpa, Honduras Accepted for Publication February 2, 2007 ABSTRACT Multiplex polymerase chain reaction (mPCR), the simultaneous amplifi- cation of two or more polymerase chain reaction (PCR) products in the same reaction tube, can be applied to the rapid detection of pathogenic microor- ganisms in food matrices. mPCR can be used to test for multiple pathogens simultaneously or to test for multiple virulence factors associated with a single pathogen. Heterogeneous pathogens such as Escherichia coli cannot only be detected by mPCR but can also be differentiated by pathotype or serotype based on amplicon profiles. Sensitive detection of pathogens in food by mPCR (1 cfu/g) can be achieved if a high concentration of target DNA with minimal contamination by PCR inhibitors can be prepared. This process generally requires enrichment, which adds 6–24 h to the assay time. Even so, pathogens that grow slowly, are difficult to isolate or are difficult to identify can be detected more rapidly by mPCR than by culture-based methods. This review covered methods and application of mPCR assays for detection of pathogens in the food production and distribution process. PRACTICAL APPLICATIONS Like uniplex PCR, mPCR requires significantly less time and is far less labor-intensive than conventional culture-based methods of testing food 5 Corresponding author: TEL: 765-496-3820; FAX: 765-494-7953; EMAIL: llperry@purdue.edu Journal of Rapid Methods & Automation in Microbiology 15 (2007) 176–198. All Rights Reserved. © 2007, The Author(s) Journal compilation © 2007, Blackwell Publishing 176