AOAC-RI ERP Book MICRO.pdf

QIAGEN

mericon

STEC Workflow Collaborative Study Protocol

May 2016

DRAFT

5.0

Candidate Method

AOAC Official Method

XXXX.XX

STEC in Selected Foods

PCR with Manual and Automated Sample Preparation

First Action XXXX

(Applicable for detection of Shiga toxin-producing

E. coli

(STEC), including serogroups O26, O45, O103,

O111, O121, O145 and O157:H7, in fresh raw ground beef and fresh raw beef trim.)

See

Table XXXX.XXA for the results of the interlaboratorystudy supporting acceptance of the

method.

Principle

QIAGEN’s

mericon

E.coliO157 Screen Plus and

mericon

E.coliSTEC O-TypePathogen Detection Assays

are multiplex PCR assays that amplify both a specific DNA target and an internal control with high

specificity. The internal control provides data regarding the presence of inhibitors in the tested samples

and the overall quality of the PCR run. Each

mericon

PCR Assay includes a PCR primer set for a pathogen-

specific target sequence, probes labeled with four distinct fluorescent dyes (

mericon

E. coliO157 Screen

Plus) or three distinct fluorescent dyes (

mericon

E. coliSTEC O-Type), positive control DNA, and all of the

reagents necessary to perform the analysis. The Multiplex PCR Master Mix included in each kit contains

QIAGEN proprietary technology, including HotStarTaq

Plus

DNA polymerase, patented multiplex PCR

technology such as Factor MP, and fast cycling technology including Q-bond.

The Rotor-Gene Q cycler produces raw data files that are further interpreted by the software using

mathematical algorithms to characterize samples. The software guides the user through each step and

provides simplicity for beginners as well as an experimental platform for advanced users.

Test Kits and Components

(

)

mericon

E. coli O157 Screen Plus

Kit.-

QIAGEN #290403, 24 tests, includes assay components,

RNase-free water, and multiplex PCR master mix.

(

)

mericon

E. coli STEC O-Type

Kit.-

QIAGEN #290233, 24 tests, includes assay components, RNase-

free water, and multiplex PCR master mix.

(

)

mericon

DNA Bacteria Kit.-

QIAGEN #69525, includes Fast Lysis buffer, product insert, and quick

start protocol.

(

)

QIAsymphonymericon® Bacteria Kit.-

QIAGEN #931156, includes reagent cartridge, piercing lid,

top elute fluid, reuse seal set, product insert, and quick start protocol.

OMAMAN-36 B : Collaborative Study Protocol

For ERP Use Only

January 2017

AOAC Research Institute

Expert Review Panel Use Only