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SUBMITTED 050113, REVISED 102513
(c)
Transfer the cluster tubes to the 80
o
C heater block and incubate for 20 min.
Note:
The
1
incubation time may be extended to a maximum of 60 min. for the purpose of managing staggered
2
assay start times.
3
(d)
Approximately 3 min. before the end of the lysis step, preheat the ANSR reaction tubes to 56
o
C
4
by placing the tubes in the incubator/reader.
Note:
The strip of tubes may be cut to provide the number
5
of tubes needed.
6
(e)
At the end of the 20 min. lysis incubation, remove and discard the caps from the reaction tubes.
7
Note
: Steps (f)-(h) should be completed without delay.
8
(f)
Using an 8-channel micropipettor and 50 µL tips with filters, carefully transfer 50 µL of the lysed
9
samples to the reaction tubes. Mix by rapidly pipetting up and down at least 10 times until the sample
10
appears homogenous in the pipette tip. Avoid excessive bubble formation by not depressing the
11
pipettor plunger beyond the first stop.
12
(g)
Place the permanent caps on the reaction tubes and close the lid of the incubator/reader.
13
(h)
Click START in the ANSR software to begin the assay.
14
(i)
The assay will complete in 10 min. and results will be displayed.
15
16
F
.
Interpretation of Results
17
The ANSR software will indicate the test results as POSITIVE, NEGATIVE, or INVALID. A positive result
18
indicates that the colony tested contains
Salmonella
spp. A negative result indicates that the colony
19
tested does not contain
Salmonella
spp. Assays producing invalid results must be repeated. The real-
20
time fluorescence curves for both the test and positive control channel can be viewed using the ANSR
21
software.
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23
Limitations
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