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DuPont™ BAX
®
System Real-Time PCR Assay for
Salmonella
: Collaborative Study
DuPont Nutrition & Health
Page 8
Orange juice (25 mL)
– Weigh 25 g test portion into 225 mL pre-warmed (35°C) universal pre-
1
enrichment broth, C(j), and swirl thoroughly. Let stand at room temperature for 55-65 minutes. Do
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not mix or adjust pH. Incubate, B(m), at 35°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
4
Orange juice (25 mL)
– Weigh 25 g test portion into 225 mL pre-warmed (41°C) BAX® System MP
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media, C(a), and swirl thoroughly. Incubate, B(m), at 39-42°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
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Nonfat dry milk (25 g)
– Pour 25 g sample slowly over the surface of 225 mL pre-warmed (35°C)
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Brilliant Green Water, C(c). Let stand at room temperature for 55-65 minutes. Do not mix or adjust
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pH. Incubate, B(m), at 35°C for 22-26 hours.
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Note: Re-growth is required for this sample type.
11
Stainless steel, ceramic tile, plastic –
Add 225 mL pre-warmed (35°C) LB, C(e), to environmental
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sponge in sample bag and swirl thoroughly. Let stand at room temperature for 55-65 minutes.
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Adjust pH to 6.8±0.2 using 1 N HCl or 1 N NaOH, C(k), if necessary. Incubate, B(m), at 35°C for 22-26
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hours.
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Stainless steel, ceramic tile, plastic –
Add 225 mL pre-warmed (35°C) BPW, C(d), to environmental
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sponge in sample bag and swirl thoroughly. Adjust pH to 6.8±0.2 using 1 N HCl or 1 N NaOH, C(k), if
17
necessary. Incubate, B(m), at 35°C for 18-24 hours.
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Cocoa (25 g)
- Weigh 25 g test portion into sterile container. Use a stomacher, B(n), to homogenize
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sample for 2 minutes with 225 mL reconstituted nonfat dry milk, C(i). Let stand at room temperature
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for 55-65 minutes, then swirl thoroughly to mix. Adjust pH to 6.8±0.2 using 1 N HCl or 1 N NaOH,
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C(k), if necessary. Add 0.45 mL 1% aqueous brilliant green dye solution, C(l), and mix well. Incubate,
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B(m), at 35°C for 22-26 hours. Transfer 10 µL enrichment to 500 µL BHI broth, C(b) before
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processing.
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White pepper (25 g)
- Weigh 25 g test portion into sterile container. Use a stomacher, B(n), to
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homogenize sample for 2 minutes with 225 mL pre-warmed (35°C) TSB, C(f). Let stand at room
26
temperature for 55-65 minutes. Adjust pH to 6.8±0.2 using 1 N HCl or 1 N NaOH, C(k), if necessary.
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Incubate, B(m), at 35°C for 22-26 hours.
28
Dry Pet Food (375 g)
– Weigh 375 g test portion into sterile container. Use a stomacher, B(n), to
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homogenize sample for 2 minutes with approximately one-third to one-half of 3375 mL pre-warmed
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(35°C) LB, C(e). Add the remainder of the pre-warmed media. Adjust pH to 6.8±0.2 using 1 N HCl or
31
1 N NaOH, C(k), if necessary. Incubate, B(m), at 35°C for 22-26 hours.
32
Note: Re-growth is required for this sample type.
33
Dry Pet Food (375 g)
– Weigh 375 g test portion into sterile container. Use a stomacher, B(n), to
34
homogenize sample for 2 minutes with approximately one-third to one-half of 3375 mL pre-warmed
35
(35°C) BPW, C(d). Add the remainder of the pre-warmed media. Adjust pH to 6.8±0.2 using 1 N HCl
36
or 1 N NaOH, C(k), if necessary. Incubate, B(m), at 35°C for 22-26 hours.
37
Note: Re-growth is required for this sample type.
38
FINAL (Version 4)