In-House Validation of Multiplex PCR for Simultaneous Detection of Shiga Toxin-Producing Escherichia coli, Listeria monocytogenes and Salmonella spp. in Raw Meats.

The aim of the study was to perform in-house validation of the developed multiplex PCR (mPCR)-based alternative method to detect Shiga toxin-producing Escherichia coli (STEC), Listeria monocytogenes (L. monocytogenes) and Salmonella spp. in raw meats following the ISO 16140-2: 2016. A comparative study of the developed mPCR against the Bacteriological Analytical Manual (BAM) method was evaluated for inclusivity and exclusivity, sensitivity and the relative level of detection (RLOD). Inclusivity levels for each target bacterium were all 100%, while exclusivity for non-target bacteria was 100%. The sensitivity of the developed mPCR was calculated based on the analysis of 72 samples of raw meat. The sensitivity of the developed mPCR was 100%. The RLOD values of the developed mPCR for STEC, L. monocytogenes and Salmonella spp. were 0.756, 1.170 and 1.000, respectively. The developed mPCR showed potential as a tool for the fast, specific and sensitive detection of the three bacteria in the raw meat industry.

Development of a food spoilage indicator for monitoring freshness of skinless chicken breast.

A colorimetric mixed-pH dye-based indicator with potential for the development of intelligent packaging, as a "chemical barcode" for real-time monitoring of skinless chicken breast spoilage, is described. Also investigated was the relationship between the numbers of microorganisms and the amount of volatile compounds. This on-package indicator contains two groups of pH-sensitive dyes, one of which is a mixture of bromothymol blue and methyl red, while the other is a mixture of bromothymol blue, bromocresol green and phenol red. Carbon dioxide (CO2) was used as a spoilage metabolite because the degree of spoilage was related to the amount of increased CO2, and which was more than the level of total volatile basic nitrogen (TVB-N) during the storage period. Characteristics of the two groups of indicator solutions were studied, as well as their response to CO2. A kinetic approach was used to correlate the response of the indicator label to the changes in skinless chicken breast spoilage. Color changes, in terms of total color difference of a mixed-pH dye-based indicator, correlated well with CO2 levels of skinless chicken breast. Trials on skinless chicken breast samples have verified that the indicator response correlates with microbial growth patterns, thus enabling real-time monitoring of spoilage either at various constant temperatures or with temperature fluctuation.

Development of a novel colorimetric indicator label for monitoring freshness of intermediate-moisture dessert spoilage.

A colorimetric mixed pH dye-based indicator with potential for the development of intelligent packaging, as a "chemical barcode" for real-time monitoring of intermediate-moisture dessert spoilage, is described. This on-package indicator contains mixed pH-sensitive dyes, bromothymol blue and methyl red, that respond through visible color change to carbon dioxide (CO(2)) as a spoilage metabolite. Both indicator solution and indicator label characteristics were studied, as well as their response to CO(2). A kinetic approach was used to correlate the response of the indicator label to the changes in intermediate-moisture dessert spoilage. Color changes, in terms of total color difference of a mixed pH dye-based indicator, correlated well with CO(2) levels of intermediate-moisture dessert. Trials on golden drop have verified that the indicator response correlates with microbial growth patterns in dessert samples, thus enabling the real-time monitoring of spoilage either at various constant temperatures or with temperature fluctuation.