171 / 1195
11
(j)
Rappaport-Vassiliadis Soya Peptone (RVS), Selenite Cystine (SC), Tetrathionate-Hajna (TT-
1
Hajna) and Tetrathionate (TT) broths and Xylose Lysine Desoxycholate (XLD), Hektoen
2
Enteric (HE), Brilliant Green Sulfa (BGS) and Bismuth Sulfite (BS) agars.–
and other
3
appropriate confirmatory media for culture confirmation.
4
5
D.
Sample Enrichment
6
7
Note: Unless otherwise specified all samples should be enriched in pre-warmed media. Unless
8
otherwise specified all samples were validated with and without a 1:50 regrowth (10 µL or
9
enriched sample in 500 µL of BHI broth) for 3 hrs at 37 ± 2
°
C before performing lysis and PCR.
10
The following matrices were validated only with a regrowth: dry pet food, peanut butter,
11
orange juice, and non-fat dry milk.
12
13
(a)
Frankfurters.
—Weigh 325 g test portion into sterile container. Add approximately one third to
14
one-half of 2925 ± 58.5 mL of sterile buffered peptone water (BPW) to each portion.
15
Homogenize for approximately 2 minutes and then add the remainder of the 2925 ml of BPW.
16
Incubate for 21 ± 3 h at 35 ± 2
°
C.
17
(b)
Ground beef
.—(1) Weigh 375 g test portion into sterile container. Add 1500 ± 75 mL of sterile
18
mTSB plus 2 mg/L novobiocin to each portion. Homogenize for approximately 2 minutes.
19
Incubate for 24 ± 2 h at 35 ± 2
°
C. (2) Weigh 25 g test portion into sterile container. Homogenize
20
sample for approximately 2 min. with 225 mL buffered peptone water, and incubate, 20-24 h at
21
35 ± 2
°
C.
22
(c)
Ground beef with soy
.—(1) Weigh 325 g test portion into sterile container. Add 975 ± 32.5 mL of
23
sterile mTSB with casaminoacids and 8 mg/L novobiocin to each portion. Homogenize for
24
approximately 2 minutes. Incubate for 22 ± 2 h at 35 ± 2
°
C. (2) Weigh 25 g test portion into
25
sterile container. Homogenize sample for 2 min. with 225 mL BPW, and incubate, 22 ± 2 h at 35
26
± 2
°
C.
27
(d)
Dry Pet Food.
—Weigh 375 g test portion into sterile container. Add approximately one third to
28
one-half of 3375 ± 67.5 mL of sterile LB or BPW to each portion. Homogenize for approximately
29
2 minutes and then add the remainder of the 2925 ml of the media. Incubate for 24 ± 2 h at 35 ±
30
2
°
C. Regrowth must be performed for this matrix.
31
(e)
Ice Cream
.—(1) Weigh 25 g test portion into sterile container. Homogenize sample with 225 mL
32
lactose broth, let stand for 60 ± 5 min at room temperature. If necessary, adjust pH to 6.8 ± 0.2
33
using 1 N HCl or 1 N NaOH, then incubate, 24 ± 2 h at 35 ± 2
°
C. (2) Weigh 25 g test portion into
34
sterile container. Homogenize sample with 225 mL BPW then incubate, 24 ± 2 h at 35 ± 2
°
C. (3)
35
Weigh 25 g test portion into sterile container. Homogenize sample with 225 mL Brilliant Green
36
Water, then incubate, 24 ± 2 h at 35 ± 2
°
C.
37
(f)
Poultry rinse
.—Combine 30 ml BPW rinsate with 30 mL of either double-strength BPW (if
38
chickens were rinsed in Butterfield’s Phosphate Buffer) or in single-strength BPW if chickens
39
were rinsed in BPW. Incubate 24 ± 2 h at 35 ± 2
°
C.
40
(g)
Peanut butter
.—(1) Weigh 25 g test portion into sterile container. Homogenize sample with 225
41
mL lactose broth, let stand for 60 ± 5 min at room temperature. If necessary, adjust pH to 6.8 ±
42
Collaborative Study Approved Protocol
Expert Review Panel Use Only