Microbiology Methods for ERP Review 3-2020

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GENE-UP SLM Matrix Extension

June 19, 2017

c) Summary of Validated Performance Claims . — The GENE ‐ UP Salmonella is equivalent to the USDA/FSIS ‐ 1 MLG 4.09 (1) for chicken carcass rinse (30 mL), raw ground chicken (25 g), beef trim (375 g), raw ground 2 pork (375 g), raw ground bison (375 g) and pasteurized liquid egg (100g). The GENE ‐ UP Salmonella is 3 equivalent to the FDA/BAM Chapter 5 Salmonella (2) for powdered egg (100 g), whey protein powder 4 (375 g), instant nonfat dried milk (375 g), dry pet food (375 g), creamy peanut butter (375 g), dark 5 chocolate (375 g) and garlic powder (375 g). 8 a) Probability of Detection (POD).— The proportion of positive analytical outcomes for a qualitative 9 method for a given matrix at a given analyte level or concentration. POD is concentration 10 dependent. Several POD measures can be calculated; POD R (reference method POD), POD C (the 11 presumptive candidate results that confirmed positive POD), POD CP (candidate method presumptive 12 result POD) and POD CC (candidate method confirmation result POD). 13 14 b) Difference of Probabilities of Detection (dPOD).— Difference of probabilities of detection is the 15 difference between any two POD values. If the confidence interval of a dPOD does not contain zero, 16 then the difference is statistically significant at the 5% level. 19 Salmonella is a motile, non ‐ spore forming, Gram ‐ negative, rod ‐ shaped bacterium in the family 20 Enterobacteriaceae . Non ‐ motile variants include Salmonella Gallinarum and Salmonella Pullorum . The 21 genus Salmonella is divided into two species that can cause illness in humans, Salmonella bongori and 22 Salmonella enterica , the latter being characterized as being the greatest public health concern. 23 24 Every year, Salmonella is estimated to cause approximately one million foodborne illnesses in the United 25 States, which correlates to approximately 64% of pathogen bacterial foodborne outbreaks. Most people 26 infected with Salmonella develop diarrhea, fever, and abdominal cramps that last 12 to 72 h after infection. 27 The illness usually lasts 4 to 7 days, and most people recover without treatment. However, for some people 28 the diarrhea may be severe enough to require hospitalization. 29 30 Any raw food of animal origin, such as meat, poultry, milk and dairy products, eggs, seafood, and some 31 fruits and vegetables may harbor Salmonella . The bacteria can survive and cause illness if the raw products 32 are not cooked to a safe minimum internal temperature. Salmonella can also cause foodborne illness 33 (salmonellosis) through cross ‐ contamination, e.g., when juices from raw meat or poultry encounter ready ‐ 34 to ‐ eat foods, such as salads. (3) 37 The GENE ‐ UP® Salmonella kit is to be used with compatible PCR strip tubes in the GENE ‐ UP® Thermocycler. 38 Each reaction vial in the GENE ‐ UP Salmonella kit contains all of the necessary components for PCR, 39 including sample ‐ specific primers and probes and an internal amplification control. The Thermocycler 40 detects fluorescence at several wavelengths (channels) to allow for multi ‐ target detection in the same 41 reaction vessel. The fluorescent signal from a sample is recorded in channel 640, while the fluorescent 42 signal for an internal amplification control is recorded in channel 705. The software automatically interprets 43 the results for the internal amplification control and determines the sample result based on the outcome of 44 the control. Both the assay for the sample and in the internal amplification control utilize dual Fluorescence 45 Resonance Energy Transfer (FRET) hybridization probes. These probes consist of two different short 46 oligonucleotides that hybridize to an internal sequence of the amplified fragment during the annealing 47 phase of the reaction cycle. The first probes for the same assay is labeled at the 3’ end with fluorescein; the 48 second probe is labeled at the 5’ end with LC Red 640. FRET occurs only after the two probes come in close 49 6 7 Definitions 17 18 General Information 35 36 Principle of the Method

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