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AOAC Official Method 2016.xxx
1
Escherichia coli
O157:H7 and
Escherichia coli
non-O157 Shiga Toxin-Producing
2
Escherichia coli
(STEC)in SelectFoodsQIAGEN
mericon
®
E. coliO157 Screen Plus and
3
mericon
®
E. coliSTEC O-Type Pathogen DetectionAssays
4
First Action 2016
5
6
Applicable to detection of
Escherichia coli
O157:H7 and
Escherichia coli
non-O157 STECin
7
fresh spinach (25g), raw ground beef (70% lean) (325g), and raw beef trim (325g).
8
9
See Tables
2016.1A - 2016.1D
for a summary of results of the inter-laboratory study.
10
See Tables
2016.2A - 2016.2D
for detailed results of the inter-laboratory study
11
12
A.
Principle
13
14
QIAGEN’s
mericon
E. coliO157 Screen Plus and
mericon
E. coliSTEC O-Type
Pathogen
15
Detection Assays
aremultiplex PCR assays that screen and confirm the presence of
E. coli
O157
16
and non-O157 STEC in food matrices. Each
mericon
PCR Assay includes a PCR primer set for a
17
pathogen-specific target sequence, probes labeled with four distinct fluorescent dyes (
mericon
E.
18
coliO157 Screen Plus) or three distinct fluorescent dyes (
mericon
E. coliSTEC O-Type), positive
19
control DNA, and all of the reagents necessary to perform the analysis. The Multiplex PCR
20
Master Mix included in each kit contains QIAGEN proprietary technology, including HotStart
21
Taq
plus
DNA Polymerase, patented multiplex PCR technology such as Factor MP, and fast
22
cycling technology including Q-bond. Extracted DNA is analyzed using the Rotor-Gene Q
®
23
cycler which uses proprietary software to detect the presence of target DNA.
24
QIAGEN, 19300 Germantown Rd., Germantown, MD 20874
25
26
B.
Apparatus and Reagents
27
Items are available from QIAGEN (Germantown, MD, 20874, USA).
28
(a)
Rotor-Gene Q5 Plex system (cat. 9001570)- For automated detection of pathogens
29
with a notebook computer containing Rotor-Gene Q Software version 2.3.1.49.
30
(b)
QIAsymphony SP/AS instrument(cat. 9001297 and cat. 9001301)– For automated
31
sample preparation, DNA extraction, and PCR preparation.
32
(c)
mericon
E. coli O157 Screen Plus Pathogen Detection Assay (24) (cat. 290403)
33
(d)
mericon
E. coli STEC O-Type Pathogen Detection Assay (24) (cat. 290233)
34
(e)
mericon
DNA Bacteria Kit (100) (cat. 69525)
35
(f)
QIAsymphony
®
mericon
DNA Bacteria Kit (360) (cat. 931156)
36
(g)
72-Well Rotor (cat. 9018903) - For holding 0.1 mL Strip Tubes and Caps.
37
(h)
Locking Ring 72-Well Rotor (cat. 9018904) - For locking 0.1 mL Strip Tubes and
38
Caps.
39
(i)
Rotor- Disc 72 Rotor (cat. 9018899) - For holding Rotor- Disc 72 (cat. 981303 or
40
981301).
41
(j)
Rotor-Disc 72 Locking Ring (cat. 9018900) - For locking Rotor-Disc
42
(k)
Insert, 2.0 mL v2, samplecarrier. (24), Qsym (cat. 9242083)
43
(l)
Cooling Adapter, EMT, v2, Qsym (cat. 920730)
44
(m)
Sample Prep Cartridges, 8-well (cat. 997002)
45
(n)
8-Rod Covers (cat. 997004)
46
(o)
Filter-Tips, 1500 µL (cat. 997024)
47
OMAMAN-36 A : Collaborative Study Manuscript
For ERP Use Only
January 2017
AO C Research Institute
Expert R view Panel Use Only