Validation of the First Marketed Multiplex Real-time PCR Rapid Test for Salmonella Enteritidis and Typhimurium Detection

Introduction: BIOTECON Diagnostics developed and validated the foodproof^® Salmonella Enteritidis & Typhimurium Detection LyoKit, a fast, sensitive and convenient assay for the detection and identification of the Salmonellaserovars SE and ST in less than 24 hours. The pre-filled reaction mix is lyophilized, and the DNA sample preparation is added directly. It is the first multiplex real-time PCR rapid test on the market, which detects SE and ST simultaneously in one single test.

Purpose: Sensitivity, specificity, and robustness were determined in combination with the StarPrep One Kit for DNA extraction.  

Methods: For exclusivity, more than 288 non-Enteritidis, non-Typhimurium Salmonella were successfully tested, including related serovars like e.g. Gallinarum.  50 non-Salmonella strains, including genetically related strains as well as strains commonly found in similar food environments, were also tested.

Results: SE was consistently detected at 10 GE/reaction or higher, while ST was detected at 5 GE/reaction or higher. Enrichments of 11 different food and PPS matrices like shell eggs, ground meat and chick dung were spiked with 10² - 10³CFU/ml, both SE and ST were successfully detected. 60 and 58 strains of SE and of ST were successfully tested, respectively.

Significance: The new assay is a safe, easy-to-use and time-saving solution with proven sensitivity, specificity, and robustness.