Validation of the Soleris E. coli method for detection and semi-quantitative determination of Escherichia coli in foods.

The performance of the Soleris E. coli method was compared with that of the ISO 7251 most probable number (MPN) and detection reference methods for Escherichia coli. The Soleris E. coli method is a growth-based, rapid, automated system composed of temperature-controlled incubation chambers and photodiode-based optical detection devices for measurement of color changes in a prepared medium vial. A dilution of the test sample homogenate is inoculated directly into the vial. Products of E. coli metabolism alter the color of the medium over time, and this change is monitored by the Soleris instrument. The test is used in a dilute-to-specification or specification monitoring manner in which the result is positive or negative around a desired cutoff (in CFU/g) determined by the dilution and volume of sample homogenate added to the vial. Alternatively, the test is used for zero tolerance determinations (e.g., absence in 25 g) by performing an off-line pre-enrichment step followed by transfer of a portion of the pre-enrichment culture to the Soleris vial. Six E. coli strains originating from food sources were inoculated individually into six food commodities: frozen green beans, Echinacea powder, cocoa powder, sweetened condensed milk, pasteurized liquid egg, and shredded mozzarella cheese. Uninoculated samples were included in each trial. The results obtained by the ISO 7251 detection method and the Soleris E. coli method were shown to be in agreement by Chi-square analysis when the presence of E. coli was determined in 25 g of sample. Results from the Soleris E. coli dilute-to-specification method and the ISO 7251 MPN method were found to be in agreement by probability of detection statistical analysis. In inclusivity testing, 52 of 53 E. coli strains were detected within 24 h. Only a non-thermoduric strain of serotype O157:H43 was not detected. In exclusivity testing, all 31 strains tested produced negative results. Results of ruggedness experiments show that accurate results can be obtained even when the operating temperature of the Soleris instrument is set beyond normal tolerances. The internal and independent laboratory studies demonstrated that the Soleris E. coli method could be successfully utilized as an alternative to the reference methods, with a significant time savings of 2 to 3 days.

Reveal Salmonella enteritidis test for detection of Salmonella enteritidis in shell eggs and environmental samples.

Reveal Salmonella Enteritidis (SE) is a lateral flow-based immunodiagnostic assay used for rapid detection of Salmonella enterica serovar Enteritidis from pooled shell eggs and environmental samples. This assay uses highly specific antibodies to accurately detect S. Enteritidis. Studies were conducted to compare the performance of this test against reference procedures for detection of S. Enteritidis from both pooled shell eggs and environmental samples. Pooled shell eggs were inoculated with low levels ofS. Enteritidis and were enriched according to the procedure prescribed by the U.S. Food and Drug Administration. Uninoculated samples were included in each trial. Reveal SE exhibited 100% sensitivity and 100% specificity in comparison to the reference method in all trials. An abbreviated 48 h/(no hold) enrichment procedure was also developed and validated for detection ofS. Enteritidis from pooled shell egg samples. This shortened enrichment procedure can be used in conjunction with the Reveal SE test and offers a significant enrichment time savings of 96 h. Chi-square analysis revealed that there was no significant difference between the abbreviated Reveal method and the reference procedure for detection ofS. Enteritidis from pooled shell egg samples. Out of 245 natural drag swabs screened internally, only three samples tested Reveal SE positive and were confirmed by the reference procedure, resulting in 100% sensitivity and 100% specificity. An external laboratory screened 147 poultry house environmental samples and obtained 35 Reveal SE confirmed positives for Reveal SE sensitivity of 100% and specificity of 90%. Inoculation trials with drag swabs resulted in 96% sensitivity and 100% specificity. Thus, these data demonstrate that Reveal SE is a highly sensitive and specific assay for the detection of S. Enteritidis from both pooled shell eggs and environmental samples.

Validation of the Soleris yeast and mold test for semiquantitative determination of yeast and mold in selected foods. Performance tested methods 040901.

The Soleris yeast and mold method, a growth-based test system with an optical detection end point, was evaluated for its ability to detect yeast and mold contamination in a wide variety of foods. The Soleris test was used in a semiquantitative manner, in which the test result is positive or negative at a threshold level determined by the dilution and volume of sample homogenate added to the Soleris test vial. By testing at two or more threshold levels, the contamination level can be estimated. The LOD of the Soleris method is 10 CFU/g when 1 mL of a 1:10 sample homogenate is added to the test vial. In these studies, the Soleris results were compared to plate counts obtained using the U.S Food and Drug Administration/Bacteriological Analytical Manual direct plating method, and agreement between the methods was calculated. Considering results from both internal and independent laboratory trials, overall agreement between the methods was 90%. Chi-square analysis showed, with few exceptions, that results of the Soleris and direct plating methods were not statistically different. Ruggedness testing was performed, and the Soleris method was found to be robust when challenged with marginally suboptimal assay conditions. Results of inclusivity testing showed that the Soleris test vial medium supports the growth of a wide variety of yeasts and molds common to foods. Results of exclusivity testing showed that bacteria do not produce positive results, even when present in the vial in relatively high initial concentrations. The Soleris method produces results in 72 h or less and thus offers considerable time savings in comparison to other commonly used yeast and mold methods.

Validation of the Reveal(®) 2.0 Group D1 Salmonella Test for Detection of Salmonella Enteritidis in Raw Shell Eggs and Poultry-Associated Matrixes.

A study was conducted to assess the performance of the Reveal(®) 2.0 Group D1 Salmonella lateral flow immunoassay for use in detection of Salmonella Enteritidis (SE) in raw shell eggs and poultry-associated matrixes, including chicken carcass rinse and poultry feed. In inclusivity testing, the Reveal 2.0 test detected all 37 strains of SE tested. The test also detected all but one of 18 non-Enteritidis somatic group D1 Salmonella serovars examined. In exclusivity testing, none of 42 strains tested was detected. The exclusivity panel included Salmonella strains of somatic groups other than D1, as well as strains of other genera of Gram-negative bacteria. In matrix testing, performance of the Reveal 2.0 test was compared to that of the U.S. Department of Agriculture, Food Safety and Inspection Service Microbiology Laboratory Guidebook reference culture procedure for chicken carcass rinse and to that of the U.S. Food and Drug Administration Bacteriological Analytical Manual for raw shell eggs and poultry feed. For all matrixes evaluated, there were no significant differences in the ability to detect SE when comparing the Reveal 2.0 method and the appropriate reference culture procedure as determined by probability of detection statistical analysis. The ability of the Reveal 2.0 test to withstand modest perturbations to normal operating parameters was examined in robustness experiments. Results showed that the test can withstand deviations in up to three operating parameters simultaneously without significantly affecting performance. Real-time stability testing of multiple lots of Reveal 2.0 devices established the shelf life of the test device at 16 months postmanufacture.

Performance comparison of the BioSys optical assay and the violet red bile agar method for detecting coliforms in food products.

Coliform counts in a variety of foods, including dairy products (raw milk, pasteurized milk, yogurt, butter, and ice cream), meats (pork sausage, ground beef, and raw chicken), raw eggs, and chocolate, were performed by the rapid automated BioSys optical assay and the conventional method with violet red bile agar (VRBA). The standard deviation (SD) among five replicate counts for the optical assay was similar to or better than that obtained with VRBA plates for all foods tested. The average SD for all foods tested was 0.21 for the optical assay and 0.30 for the VRBA plates. At very low concentrations of coliforms (1 to 10 CFU/ml for liquid products and 10 to 100 CFU/g for solid samples), the average SDs were 0.26 and 0.47, respectively. The optical assay was less susceptible to interference by noncoliform organisms. In naturally contaminated samples, bacteria such as Serratia liquefaciens, Pantoea spp., Vibrio fluvialis, Aeromonas hydrophilia, and Pseudomonas spp. formed typical colonies in VRBA, resulting in false-positive results or a need to verify colonies in brilliant green lactose broth. The optical assay appeared to be more selective than the VRBA conventional method, detecting fewer noncoliforms. There was close agreement in test results between the two methods, as indicated by correlation coefficients of 0.92 to 0.99 obtained for the regression analysis of the two methods. In most cases both methods distinguished accurately between positive samples containing coliforms and negative controls. All products tested using the automated BioSys Optical Assay for coliforms yielded results more quickly (typically 10 to 12 h) than did those tested with the conventional VRBA method (24 to 72 h with confirmation).

Matrix Extension Study: Validation of the ANSR® Salmonella Method for Detection of Salmonella spp. in Pasteurized Egg Products.

A matrix extension study was conducted to validate the ANSR(®) Salmonella method for use with pasteurized egg products. Four diverse egg product types were tested by the ANSR method and the U.S. Department of Agriculture's Microbiology Laboratory Guidebook reference culture procedure. There were no significant differences in the number of positive test portions by the two methods for any of the products examined. Independent laboratory testing of pasteurized liquid egg also found no significant difference in performance between the ANSR and reference culture procedures. There were no false-positive results obtained in either internal or independent laboratory testing. Inclusivity testing using a new enrichment medium specifically designed for use with pasteurized egg products produced 112 positive results out of 113 Salmonella spp. strains tested, with only a single strain of S. Weslaco testing negative by the ANSR assay. Exclusivity testing of 38 non-salmonellae produced all negative ANSR results. It is concluded that ANSR Salmonella is a reliable, sensitive, and specific method for detection of Salmonella spp. in pasteurized egg products.