Microbiology Methods for ERP Review 3-2020

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Homogenize by hand massaging for 2 min ± 30 s and incubate at 41.5 ± 1°C for 20–24 h. 1 ( f ) Honey Mustard Onion Seasoning and Flavored Ranch Seasoning (375 g test portions) 2 Add 3,375 mL of Solus mBPW Medium supplemented with Solus Salmonella supplement [2.22 mL 3 Solus Salmonella supplement per 1,000 mL Solus mBPW] to a 375 g test portion in the sampling bag. 4 Homogenize by hand massaging for 2 min ± 30 s and incubate at 41.5 ± 1°C for 20–24 h. 5 ( g ) Instant Non ‐ Fat Dry Milk Powder (375 g test portions) 6 Pre ‐ warm 1,125 mL of Solus BPW (ISO) Medium supplemented with Solus Salmonella supplement 7 [4.44 mL Solus Salmonella supplement per 1,000 mL Solus BPW (ISO)] to 37 ± 1°C for at least 2 h prior to 8 its addition to a 375g test portion in the sampling bag. Homogenize by hand massaging for 2 min ± 30 s 9 and incubate at 41.5 ± 1°C for 21–22 h. 10 ( h ) Stainless Steel environmental surface 11 For 4” x 4” surface area, sample surface with sterile sampling sponge device (e.g. World Bioproducts 12 EZ Reach Sponge Sampler with HiCap neutralizing broth or Hygiena equivalent). Follow sampling device 13 manufacturers’ instructions for correct use, storage and transport. Pre ‐ warm 100 mL of Solus BPW (ISO) 14 Medium supplemented with Solus Salmonella supplement [4.44 mL Solus Salmonella supplement per 15 1,000 mL Solus BPW (ISO) to 37 ± 1°C for at least 2 h prior to its addition to the sampled sponge in the 16 sampling bag. Homogenize by hand massaging for 2 min ± 30 s and incubate at 41.5 ± 1°C for 16–20 h. 17 ( i ) Plastic environmental surface 18 For 1” x 1” surface area, sample surface with sterile sampling swab (e.g. sponge or foam pre ‐ 19 moistened with Letheen broth). Follow sampling device manufacturers’ instructions for correct use, 20 storage and transport. Pre ‐ warm 10 mL of Solus BPW (ISO) Medium supplemented with Solus 21 Salmonella supplement [4.44 mL Solus Salmonella supplement per 1,000 mL Solus BPW (ISO)] to 37 ± 22 1°C for at least 2 h prior to its addition to the sampled swab in the sampling tube. Homogenize by vortex 23 mixing for 2 min and incubated at 41.5 ± 1°C for 16–20 h. 24 ( j ) Following the incubation, gently agitate each bag or tube and transfer a 1 mL aliquot to a 25 polypropylene test tube. Heat tubes at a temperature of 85–100°C for 15 – 20 min. 26 ( k ) After heat treatment, cool the samples to room temperature (15–25°C). This may be 27 accelerated by placing the test tubes in cold water for 5 min. 28 ( l ) Where appropriate use frit filters with denatured matrices that contain high concentrations of 29 particulates to minimize sample handling errors. 30 ( m ) Keep the remaining non ‐ heat ‐ treated samples for verification until ELISA results are obtained. 31 These samples can be stored at 41.5 ± 1°C if the ELISA test is to be carried out within 2 h or at 2–8°C for 32 up to 72 h if not.

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G. Solus One Salmonella (Manual Method)

( a ) Remove the kit from storage at 2–8°C an hour before use to allow the components to reach 37 room temperature (15–25°C). Determine the number of wells required for the test. Take the required 38 number of strips from the pouch and fit them to the framer provided. Note: Unused strips should be 39 returned to the pouch and stored at 2–8°C. 40 ( b ) Prepare the Washing Buffer by adding the contents of a single Washing Buffer Activator 41 sachet and Washing Buffer bottle (60 mL) to 1440 mL or a single Washing Buffer Activator sachet and 42

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