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REVISED 121213
days). Collaborators were provided with a “Sample Receipt Form”, to be completed and returned to the
1
Study Director by email or fax, acknowledging that the samples were received in good condition.
2
Analysis of Isolates
3
To initiate the analysis, collaborators streaked each of the 12 bacterial isolates to each of the 7 agar
4
media, streaking for isolated colonies. Collaborators were provided with a sample randomization
5
scheme by the Study Director and were instructed to blind-code each strain-agar medium combination
6
with a unique number 1-84. This was performed by “Operator 1”, who would have no involvement in
7
the actual ANSR analyses. Plates were incubated for 24 ± 2 h at 35 ± 1
o
C and examined for the presence
8
of isolated colonies. Plates without isolated colonies were re-incubated for an additional 18-24 h. Plates
9
containing distinct isolated colonies after 24 h were stored at 2-8
o
C. After a maximum of 48 h
10
incubation, plates without growth or isolated colonies were noted as such on the Data Recording Form
11
and analysis continued. Operator 1 then picked a single colony from each plate, including the
12
refrigerated plates, using an inoculating loop or needle, and resuspended the colony in 0.5 mL
13
phosphate-buffered saline (PBS). The coded tubes were transferred to “Operator 2”, who then
14
performed the ANSR analyses. ANSR testing was performed in blocks of up to 16 samples, starting with
15
sample number 1 and continuing through sample number 84. Completed “Data Recording Forms” were
16
returned to the Study Director by email or fax. ANSR assay raw data was provided to the Study Director
17
by email as .json files. This raw data included the real-time fluorescence curves for each assay
18
performed.
19
20
AOAC Official Method
XXXX.XX21
ANSR
Salmonella
Confirmation Test
22
for Identification of
Salmonella
spp. from Colony Picks
23
First Action XXXX
24
(Applicable to the identification of
Salmonella
spp.
25
from colony picks from selective/differential agar media
26
[bismuth sulfite agar, brilliant green sulfa agar, double-modified lysine iron agar,
27
Hektoen enteric agar, tryptic soy agar, xylose lysine deoxycholate agar, xylose lysine tergitol agar (XLT-
28
4)])
29
30
4