Table of Contents Table of Contents
Previous Page  164 / 596 Next Page
Information
Show Menu
Previous Page 164 / 596 Next Page
Page Background

Environmental samples

1

Sample collection devices can be a sponge hydrated with a neutralizing solution to inactivate the

2

effects of the sanitizers. 3M recommends the use of a biocide-free cellulose sponge. Neutralizing

3

solution can be Dey-Engley (D/E) Neutralizing Broth or Letheen Broth. It is recommended to

4

sanitize the area after sampling.

5

6

The recommended size of the sampling area for verifying the presence or absence of the

7

pathogen on the surface is at least 100 cm

2

(10 cm x 10 cm or 4”x4”). When sampling with a

8

sponge, cover the entire area going in two directions (left to right then up and down) or collect

9

environmental samples following your current sampling protocol or according to the FDA BAM,

10

USDA FSIS MLG or ISO 18593 [7] guidelines.

11

12

1.

Allow the Demi-Fraser Broth enrichment medium (includes FAC) to equilibrate to ambient

13

laboratory temperature.

14

2.

Aseptically combine the enrichment medium and sample according to Table 1.

15

3.

Homogenize thoroughly by blending, stomaching, or hand mixing for 2 ±0.2 minutes.

16

Incubate at 37 ±1°C for 24-30 hours.

17

4.

Invert room temperature (20-25

o

C) lysis tubes to mix. Proceed to next step within 4 hours.

18

20 µL aliquot of each enriched sample are is transferred to separate lysis tubes using a new

19

pipette tip after each sample transfer. Place uncovered samples on a dry double block heater r

20

for 15 ± 1 minutes at 100 ± 1

o

C. Following the heat lysis transfer samples into the chill

21

block insert and placed onto a sterilized lab bench and allow to cool at 18-28 °C for 5-10

22

minutes.

23

24

5.

Add 20 µL of each lysed sample and control to separate reagent tubes, and mix by pipetting

25

up and down five times. Analyze a matrix control tube with the samples for each matrix to

26

verify that no interference with the assay was caused by the matrix. Use sterile Demi Fraser

27

Broth with FAC for the Negative Control (NC).

28

6.

Transfer a 20 µL aliquot to the NC and the Reagent Control (RC) tubes. Add a 20 µL aliquot

29

of a randomly picked sample to the matrix control tube, mixed, and recapped. Using the 3M

30

software, follow prompts to identify samples and controls. Load all samples into the Speed

31

Loader Tray (SLT), place into the Molecular Detection System, and initiate the 3M

MDA 2

32

-

Listeria

assay. Results are obtained within 75 minutes.

33

34

Interpretation and Test Result Report

35

An algorithm interprets the light output curve resulting from the detection of the nucleic acid

36

amplification. Results are analyzed automatically by the software and are color-coded based on

37

the result. A Positive or Negative result is determined by analysis of a number of unique curve

38

parameters. Presumptive positive results are reported in real-time while Negative and Inspect

39

results will be displayed after the run is completed.

40

41

NOTE:

Even a negative sample will not give a zero reading as the system and 3M Molecular

42

Detection Assay 2 -

Listeria

amplification reagents have a “background” relative light unit

43

(RLU) reading.

44

45

In the rare event of any unusual light output, the algorithm labels this as “Inspect.” 3M

46

recommends the user to repeat the assay for any Inspect samples. If the result continues to be

47

AOAC Research In titute

Expert Review Panel Use Only

OMAMAN-29 D/ PTM Validation Report 111501

OMA ERP - June 2016

ERP Use Only