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GENE-TRAK®
Listeria monocytogenes Assay
Results
Positive Negative Positive Negative
Control Control Sample Sample
Product Specifications
Sensitivity: 1-5 CFU/25 g
Testing time: 2 hours (after 40-48 hour enrichment)
Tests per kit: Up to 98
Intended Use Approvals: BAM Chapter 10.071
GENE-TRAK Listeria monocytogenes Assay provides for the detection USDA/FSIS MLG2
of L. monocytogenes in food and environmental samples.
Materials Required But Not Provided
The Test Available from Neogen*
The DNA hybridization test employs L. monocytogenes-specific DNA 1. GENE-TRAK photometer set
probes and a colorimetric detection system for the detection of L. 2. Test tube racks
monocytogenes species in food and environmental samples following 3. Dipstick holders
broth culture enrichment. A sample is considered negative for the 4. Wash basins with lids
presence of L. monocytogenes if the absorbance value obtained is less 5. Dilution chart
than or equal to the established cutoff value for the assay. A sample is 6. Pipettes (1 mL and 10 mL)
considered presumptively positive for the presence of L. monocytogenes 7. Incubator at 35°C
if the absorbance value obtained is greater than the established cutoff 8. 12 x 75 mm glass test tubes
value of the assay. 9. Minute timer
10. Micropipette and tips, 100 – 1000 µL range (optional)
Test Procedure 11. Modified LCA
1. Enrich food samples. Not available from Neogen
2. Add 0.5 mL of enriched samples, negative control and positive 1. Blender or homogenizer
control into separate 12 X 75 mm glass tubes. 2. Culture bottles for sample enrichments
3. Add 0.1 mL pretreatment reagent and incubate at 37°C for 15 3. Phosphate-buffered Listeria enrichment broth
minutes. 4. Phosphate-buffered saline
4. Add 0.1 mL lysis reagent to each sample tube. Shake for 5 seconds 5. UVM-1 broth
and incubate at 37°C for 15 minutes. 6. UVM-2 broth
5. Add 0.1 mL L. monocytogenes probe solution to each tube. 7. Modified UVM-2 broth
6. Place dipsticks into sample tubes. Raise and lower dipsticks 5 times 8. Sterile cotton swabs
to mix and incubate at 65°C for 1 hour. 9. Water baths or heater blocks capable of maintaining 37°C ± 1°
7. Wash dipsticks two times each for 1 minute at 65°C and 1 minute and 65°C ± 1°
at room temperature. 10. Graduated cylinder
8. Transfer the dipsticks to tubes containing 0.75 mL of enzyme 11. Absorbent paper
conjugate and incubate at room temperature for 20 minutes. 12. Sterile tubes with caps for 1 mL volume
9. Wash dipsticks two times for 1 minute each at room temperature.
U.S. FDA. 1998. Bacteriological Analytical Manual, 8th ed., Rev. A.
1
10. Transfer dipsticks to tubes containing 0.75 mL of substrate- USDA/FSIS Microbiology Laboratory Guidebook, 3rd ed., Rev. 2, Nov. 8, 1999.
2
chromogen and incubate at room temperature for 30 minutes. Chapter 8.
11. Remove dipsticks and add 0.25 mL stop solution. * See Equipment and Accessories, pages 91–95
12. Read results using a GENE-TRAK photometer.
Ordering Information
Prod.# Product description
6707 GENE-TRAK Listeria monocytogenes Assay - up to 98
tests
44 Sales & Service: 800/234-5333 (USA/Canada) or 517/372-9200 • Fax: 517/372-2006 • www.neogen.com
