The AccuPoint Advanced ATP Hygeine Monitoring System was validated by an AOAC International Performance Tested MethodSM on the detection of ATP from stainless steel surfaces. Neogen Corp.'s system is a lightweight, hand-held diagnostic tool used to validate and verify a hygiene program's effectiveness by detecting organic residues remaining on surfaces and in liquids after cleaning. The system is composed of three primary components: an electronic luminometer, fully self-contained single-use samplers, and software. The system is designed to detect adenosine triphosphate (ATP) at set thresholds and to report the measurement in relative light units (RLU). These thresholds are established by a facility to reflect effective cleaning practices. The instrument compares the measured level of ATP with the established threshold and reports the results as pass, marginal, or fail. A linear dose-response in RLU was observed with pure analyte. In the matrix and microbial studies, detection levels varied depending on the matrix and microorganism tested. Independent laboratory trials confirmed pure analyte and matrix observations. Specificity testing of similar, yet different, compounds resulted in 0 RLU for all except 2'-deoxyadenosine 5'-triphosphate sodium salt, which showed markedly reduced reactivity when compared with ATP. Also, interference by these compounds was negligible. When disinfectant residues were evaluated for their effect on the test, cleaners increased RLU output to varying degrees. Stability testing showed consistent results between three independently manufactured lots and stable results through the 9 month shelf-life. Additionally, when three readers were compared using electronic light-emitting diodes as the light source, instrument variability was low (<3%). Robustness testing results provided evidence that temperature affects test performance more than shaking time, and sampler performance improves as the temperature increases to room temperature. These results provided evidence that the AccuPoint Advanced ATP Hygiene Monitoring System produces consistent and reliable data for the evaluation of sanitation program effectiveness on stainless steel surfaces in food processing and food service facilities.
Background Many food recalls are related to the presence of undeclared allergens and microorganisms in food products. To reduce these occurrences, portable diagnostic assay kits are available to quantitate mycotoxins, detect allergens and gluten in foods and on environmental surfaces and for sanitation monitoring. Objective This article reviews diagnostic kits that can detect sources of contamination in food and ingredients as well as on surfaces and clean in place (CIP) rinses. Methods Mycotoxins and gluten were detected using lateral flow diagnostic (LFD) assays. Sanitation monitoring of surfaces was completed using a chemiluminescent assay to detect ATP and another assay to detect protein. Results Gluten was detected at 10 ppm in spiked commodities and on wet and dry surfaces at 2.5 µg/100cm2. Deoxynivalenol was quantitated in Dry Distillers Grains plus Solubles (DDGs) and mean results were within two standard deviations (SD) of those determined by high performance liquid chromatography (HPLC). The chemiluminescent assay had a limit of detection (LOD) of 6 femtomoles of ATP and was able to detect a 1:10,000 dilution of orange juice from surfaces. The protein assay detected 5 µg of BSA directly applied to the sampler, 100 µg of BSA on surfaces, and detected 1:10 dilutions of Greek yogurt and raw beef from surfaces. Conclusions Portable diagnostic kits evaluated in this work provided accurate, rapid and sensitive results for detection of mycotoxins, gluten, proteins, and ATP. These methods can be used in facilities with minimal training and provided results that are important to ensure food safety. Highlights Portable methods to detect gluten, mycotoxins, proteins and ATP are presented.
Reveal E. coli 2.0 is a new lateral-flow immunodiagnostic test for detection of E. coli O157:H7 and O157:NM in raw beef trim and ground beef. Compared with the original Reveal E. coli O157:H7 assay, the new test utilizes a unique antibody combination resulting in improved test specificity. The device architecture and test procedure have also been modified, and a single enrichment protocol was developed which allows the test to be performed at any point during an enrichment period of 12 to 20 h. Results of inclusivity and exclusivity testing showed that the test is specific for E. coli serotypes O157:H7 and O157:NM, with the exception of two strains of O157:H38 and one strain of O157:H43 which produced positive reactions. In internal and independent laboratory trials comparing the Reveal 2.0 method to the U.S. Department of Agriculture-Food Safety and Inspection Service reference culture procedure for detection of E. coli O157:H7 in 65 and 375 g raw beef trim and ground beef samples, there were no statistically significant differences in method performance with the exception of a single internal trial with 375 g ground beef samples in which the Reveal method produced significantly more positive results. There were no unconfirmed positive results by the Reveal assay, for specificity of 100%. Results of ruggedness testing showed that the Reveal test produces accurate results even with substantial deviation in sample volume or device incubation time or temperature. However, addition of the promoter reagent to the test sample prior to introducing the test device is essential to proper test performance.
Reveal Salmonella 2.0 is an improved version of the original Reveal Salmonella lateral flow immunoassay and is applicable to the detection of Salmonella enterica serogroups A–E in a variety of food and environmental samples. A Performance Tested MethodSM validation study was conducted to compare performance of the Reveal 2.0 method with that of the U.S. Department of Agriculture-Food Safety and Inspection Service or U.S. Food and Drug Administration/Bacteriological Analytical Manual reference culture methods for detection of Salmonella spp. in chicken carcass rinse, raw ground turkey, raw ground beef, hot dogs, raw shrimp, a ready-to-eat meal product, dry pet food, ice cream, spinach, cantaloupe, peanut butter, stainless steel surface, and sprout irrigation water. In a total of 17 trials performed internally and four trials performed in an independent laboratory, there were no statistically significant differences in performance of the Reveal 2.0 and reference culture procedures as determined by Chi-square analysis, with the exception of one trial with stainless steel surface and one trial with sprout irrigation water where there were significantly more positive results by the Reveal 2.0 method. Considering all data generated in testing food samples using enrichment procedures specifically designed for the Reveal method, overall sensitivity of the Reveal method relative to the reference culture methods was 99%. In testing environmental samples, sensitivity of the Reveal method relative to the reference culture method was 164%. For select foods, use of the Reveal test in conjunction with reference method enrichment resulted in overall sensitivity of 92%. There were no unconfirmed positive results on uninoculated control samples in any trials for specificity of 100%. In inclusivity testing, 102 different Salmonella serovars belonging to serogroups A–E were tested and 99 were consistently positive in the Reveal test. In exclusivity testing of 33 strains of non-salmonellae representing 14 genera, 32 were negative when tested with Reveal following nonselective enrichment, and the remaining strain was found to be substantially inhibited by the enrichment media used with the Reveal method. Results of ruggedness testing showed that the Reveal test produces accurate results even with substantial deviation in sample volume or device development time.
ANSR® Listeria was previously certified as Performance Tested Method(SM) 101202 for detection of Listeria spp. on selected environmental surfaces. This study proposes a matrix extension to the method for detection of Listeria spp. in selected food matrixes. The method is an isothermal nucleic acid amplification assay based on the nicking enzyme amplification reaction technology. Following single-step sample enrichment for 16-24 h, the assay is completed in less than 50 min, requiring only simple instrumentation. Inclusivity testing was performed using a panel of 51 strains of Listeria spp., representing the species L. grayi, L. innocua, L. ivanovii, L. monocytogenes, L. seeligeri, and L. welshimeri. All strains tested were detected by the ANSR assay. Exclusivity testing of 30 strains representing non-Listeria Gram-positive bacteria yielded no evidence of cross-reactivity. Performance of the ANSR method for detection of Listeria spp. was compared to that of reference culture procedures for pasteurized liquid egg, pasteurized 2% milk, Mexican-style cheese, ice cream, smoked salmon, lettuce, cantaloupe, and guacamole. Data obtained in these unpaired studies and analyzed using a probability of detection model demonstrated that there were no statistically significant differences in results between the ANSR and reference culture methods, except for milk at 16 h and cantaloupe. In milk and smoked salmon, ANSR sensitivity was low at 16 h and therefore the recommended incubation time is 24 h. In cantaloupe, ANSR was found to be more sensitive than the reference culture method at both 16 and 24 h in independent laboratory testing. The ANSR Listeria method can be used as an accurate, rapid, and simple alternative to standard culture methods for detection of Listeria spp. in selected food types.
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