Comparison between the Real-Time PCR and Crystal Diagnostic Xpress Immunoassay Methods for Detecting Salmonella and Shiga Toxin–Producing Escherichia coli in the Air of Beef Slaughter Establishments

Abstract

ABSTRACT The aim of this study was to compare the effectiveness of a quantitative real-time PCR (qPCR) molecular method and the Crystal Diagnostic Xpress (CDx) immunoassay for detecting Salmonella and Shiga toxin–producing Escherichia coli (STEC) in air samples collected from abattoirs in Texas. The 70 air samples were collected from two small and two large meat processing plants in the spring and summer with a wetted wall cyclone air sampler. The samples were divided equally into two parts: one part was used for the qPCR assay, and the other part was enriched for 18 and 36 h and evaluated with the CDx immunoassay. All samples for which positive results were obtained were confirmed by plating and by biochemical and serological tests as recommended by AOAC International to verify results of rapid methods. With the qPCR and CDx assays and 36 h of enrichment, 37.5 and 57.1% of the samples, respectively, were positive for Salmonella (P < 0.05) and 65.0 and 60.7%, respectively, were positive for STEC (P > 0.05). Air samples required longer enrichment for the CDx immunoassay than recommended by the manufacturer for food samples. Recovery of Salmonella and STEC increased 16 and 47%, respectively, when enrichment was extended from 18 to 36 h. The prevalence of Salmonella and STEC obtained with both methods was affected by the size of the processing plant and the processing stage. Detection rates for samples from larger plants were higher for both pathogens. Significantly higher prevalence was obtained for samples from the stunning and dehiding areas than for those from the fabrication rooms and chillers. Salmonella detection was higher with the CDx assay than with the qPCR assay, but no differences were found for the detection of STEC by the qPCR and CDx assays. These results highlight the importance of method adjustments when testing matrices other than foods. More research is needed to understand the dynamics of pathogen dispersal in aerosols and how this affects the effectiveness of current rapid detection methods. HIGHLIGHTS