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Foodborne Bacteria www.neogen.com/foodsafetyorder.htm

GENE-TRAK®
Generic E. coli Assay

Results

Positive Negative Positive Negative


Control Control Sample Sample

Product Specifications
Sensitivity: 1-5 CFU/25 g
Testing time: 2 hours (after 24-48 hour
enrichment)
Tests per kit: Up to 98
Intended Use
GENE-TRAK E. coli Assay provides for the detection of E. coli in food. Materials Required But Not Provided
Available from Neogen*
The Test 1. GENE-TRAK photometer set
The DNA hybridization test employs E. coli-specific DNA probes and a 2. Pipettes (1 mL and 10 mL)
colorimetric detection system for the detection of E. coli in food samples 3. Incubator at 35°C
following broth culture enrichment. The method is a semi-quantitative 4. 12 x 75 mm glass test tubes
test to detect the presence of E. coli at contamination levels greater than 5. Minute timer
3 cells per gram in the original food sample. A sample is considered 6. Micropipette and tips, 100-1000 mL range (optional)
non-reactive for the presence of E. coli if the absorbance value obtained 7. Test tube racks
is less than or equal to the established cutoff value for the assay. A sample 8. Dipstick holders
is considered reactive for the presence of E. coli if the absorbance value 9. Wash basins w/lids
obtained is greater than the established cutoff value of the assay. 10. Dilution chart (for enzyme conjugate)

Test Procedure Not available from Neogen


1. Blender or homogenizer
1. Enrich food sample.
2. Culture bottles to hold 500 mL sample volume
2. Add 0.5 mL of enriched samples to tubes and 0.5 mL of posi-
3. Covered water bath at 65°C
tive and negative controls to separately labeled 12 x 75 mm glass
4. Graduated cylinder
tubes.
5. Absorbent paper
3. Add 0.1 mL lysis reagent to each sample tube. Shake for 5 seconds
6. Sterile tubes with caps for 1 mL volume
and then incubate at room temperature for 5 minutes.
7. Butterfield’s phosphate-buffered dilution water
4. Add 0.2 mL E. coli probe solution to each tube. Shake the rack
8. Lauryl Sulfate Tryptose Broth
of tubes by hand for 5 seconds. Place the rack of tubes in 65°C
9. 42°C (24 hr enrichment protocol) water bath
water bath and incubate for 15 minutes.
5. Place dipsticks into sample tubes. Raise and lower dipsticks 5 * See Equipment and Accessories, pages 91–95
times to mix and incubate at 65°C for 1 hour.
6. Wash dipsticks for 1 minute at room temperature.
7. Transfer the dipsticks to tubes containing 0.75 mL of enzyme
conjugate and incubate at room temperature for 20 minutes.
8. Wash the dipsticks two times for 1 minute each at room
temperature.
9. Transfer the dipsticks to tubes containing 0.75 mL of substrate-
chromogen and incubate at room temperature for 20 minutes.
10. Remove dipsticks and add 0.25 mL stop solution.
11. Read results using a GENE-TRAK photometer.
See package insert for complete instructions. Ordering Information
Prod.# Product description
6710 GENE-TRAK Escherichia coli Assay – up to 98 tests

Sales & Service: 800/234-5333 (USA/Canada) or 517/372-9200 • Fax: 517/372-2006 • www.neogen.com 55