Evaluation of Applied Biosystems MicroSEQ^® Real-Time PCR System for Detection of Listeria spp. in Food and Environmental Samples

Author： Petrauskene Olga V. Cao Yanxiang Zoder Patrick Wong Lily Y. Balachandran Priya Furtado Manohar R. Tebbs Robert S.

Publisher： AOAC International

ISSN： 1944-7922

Source： Journal of AOAC International, Vol.95, Iss.4, 2012-07, pp. : 1074-1083

Disclaimer: Any content in publications that violate the sovereignty, the constitution or regulations of the PRC is not accepted or approved by CNPIEC.

Previous Menu Next

Abstract

A complete system for real-time PCR detection of Listeria species was validated in five food matrixes and five environmental surfaces, namely, hot dogs, roast beef, lox (smoked salmon), pasteurized whole cow's milk, dry infant formula, stainless steel, plastic cutting board, ceramic tile, rubber sheets, and sealed concrete. The system consists of the MicroSEQ^® Listeria spp. Detection Kit, two sample preparation kits (PrepSEQ^® Nucleic Acid Extraction Kit and PrepSEQ Rapid Spin Sample Preparation Kit), the Applied Biosystems 7500 Fast Real-Time PCR instrument, and the RapidFinder™ Express v1.1 Software for data analysis. The test method was compared to the ISO 11290-1 reference method using an unpaired study design. The MicroSEQ Listeria spp. Detection Kit and the ISO 11290-1 reference method showed equivalent detection based on Chi-square analysis for all matrixes except hot dogs. For hot dogs, the MicroSEQ method detected more positives than the reference method for the low- and high-level inoculations, with all of the presumptive positives confirmed by the reference method. An independent validation study confirmed these findings on lox and stainless steel surface. The MicroSEQ kit detected all 50 Listeria strains tested and none of the 31 nontarget bacteria strains.