Real-Time TaqMan PCR for Rapid Detection and Quantification of Coliforms in Chilled Meat

Abstract

Coliforms are a major group of bacteria associated with spoilage of refrigerated pork. It is necessary to develop an efficient and fast method to detect total coliforms in chilled pork because the traditional methods are laborious and time-consuming. In this study, a quantitative real-time polymerase chain reaction (qRT-PCR) targeting the lacZ gene was developed for rapid enumeration of coliforms from chilled pork. Of the 106 strains tested, 35 coliform strains and one Shigella sonnei strain were correctly identified compared with 70 control strains which failed to amplify. Detection sensitivity was 112 CFU/g. The assay was able to detect and accurately quantify the total coliforms in chilled meat within 2 h without preenrichment. The results indicate that the real-time PCR is an efficient and time-saving method that could be adopted by the meat industry to detect coliforms to replace the traditional most probable number and the rapid coliform count plate methods.