AOAC-RI ERP Book MICRO.pdf

QIAGEN

mericon

STEC Workflow Collaborative Study Protocol

May 2016

DRAFT

Table 1. Study Schedule (Specific Dates TBD)

Collaborator

Activity

Receipt of

Media and

Reagents

Receipt of Test

Kits and

Materials

Receipt of Test

Portions Set

Initiation of

Analyses

Submission of

Data Report

Form

Date

Week 1,

Wednesday

Week 1,

Wednesday

Week 1,

Friday

Week 2,

Monday

Week 4,

Monday

Safety Precautions

mericon E. coli O157 Screen Plus Kit, mericon E. coli STEC O-Type

Kit,

and mericon DNA Bacteria Kit.-

All chemicals should be considered potentially hazardous. When working with chemicals, always

wear a suitable lab coat and disposable gloves. Product usage should follow good laboratory

practices.

QIAsymphonymericonBacteria Kit.-

The buffers in the reagent cartridge contain guanidine salts, which

can form highly reactive compounds when combined with bleach. If liquid containing these buffers is

spilled, clean with a suitable laboratory detergent and water. If the spilled liquid contains potentially

infectious agents, clean the affected area first with laboratory detergent and water, and then with 1%

(v/v) sodium hypochlorite solution. Reagents in this kit are highly flammable and are harmful by

inhalation, skin contact and swallowing.

Real-Time PCR System

.-Improper use of the Rotor-Gene Q may cause personal injuries or damage to

the instrument. The Rotor-Gene Q must only be operated by qualified personnel who have been

appropriately trained. Servicing of the Rotor-Gene Q must only be performed by QIAGEN Field

Service Specialists.

Enrichment.-

Shiga-toxin producing

E. coli

is a Biosafety Level 2 organism. Biological samples such as

enrichments have the potential to transmit infectious diseases. Follow all applicable local,

state/provincial, and/or national regulations on disposal of biological wastes. Wear appropriate

protective equipment, which includes but is not limited to protective eyewear, face shield,

clothing/lab coat, and gloves. All work should be conducted in properly equipped facilities utilizing

the appropriate safety equipment (for example, physical containment devices). Individuals should

be trained in accordance with applicable regulatory and company/institution requirements before

working with potentially infectious materials. All enrichment broths should be sterilized following

any culture-based confirmatory steps.

Test Portion Analysis

On Monday, all collaborators shall initiate the enrichment of test portions according to the

mericon

and

reference methods. A copy of the detailed methods will be provided for you. Note that the ground beef

is a paired study, so the same enrichments are used for the

mericon

and MLG methods. Refer to the

Study Flowchart in Appendix 8.3 for an overview of the matrix study and methods. All test portions are

pre-weighed. An additional uninoculatedtest portion is labeled for aerobic plate count (APC)

determination. Initiate the APC determination using the FDA BAM method on the same day as the study

test portions.

OMAMAN-36 B : Collaborative Study Protocol

For ERP Use Only

January 2017

AOAC Research Institute

Expert Review Panel Use Only