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GeneQuence™
E. coli O157:H7

Results

Product Specifications
Sensitivity: 1 CFU/25 grams
Testing time: 1 hour, 20 minutes
(after enrichment of 8 or 20 hours)
Intended Use Tests per kit: Up to 94
Neogen’s GeneQuence E. coli O157: H7 is intended for the screening
of meat products for the presence of E. coli O157:H7. Materials Required But Not Provided
Available from Neogen*
The Test 1. Incubator capable of maintaining 35-37°C
GeneQuence E. coli O157:H7 is a double antibody (sandwich) enzyme- 2. Reveal 8 Hour E. coli O157:H7 enrichment broth, or Reveal
linked immunosorbent assay (ELISA) utilizing specific anti-E. coli 20 Hour E. coli O157:H7 enrichment broth
O157:H7 antibodies coated to microwells. The controls and samples 3. Microwell plate or strip reader capable of reading at 450 nm
are added to the antibody wells. After incubation, the wells are washed 4. Squeeze wash bottle with fine tip, or microplate/microstrip washer
with a wash buffer and conjugate is added. After a second incubation 5. 100 µL pipettor and tips
and wash, substrate is added, which develops a blue color. A final in- 6. 13 x 100 mm glass test tubes with caps
cubation is followed by the addition of the stop solution, which turns 7. Heating block or water bath capable of maintaining 100°C
the blue color to yellow. Results are read using a microwell plate or strip 8. Weigh scale
reader by comparing the optical densities of the samples to the optical
densities of the negative and positive controls. Not available from Neogen
1. Distilled water
The Procedure * See Equipment and Accessories, pages 91–95
1. Enrich food samples.
2. Transfer 5 mL of each enriched sample to a glass test tube.
3. Heat kill samples in each tube by placing into a boiling water
bath for 10 minutes.
4. Allow samples to cool to room temperature.
5. Add 100 µL of negative control to first microwell.
6. Add 100 µL of positive control to second microwell. Ordering Information
7. Add 100 µL of enriched sample to well #3, and so on.
8. Incubate for 20 minutes at room temperature. Prod.# Product description
9. Wash 5 times with wash solution. Dry wells by tapping out 6716 GeneQuence E. coli O157:H7 – Contains 96 antibody-
wells on a lint-free towel. coated wells for up to 94 tests
10. Add 100 µL of enzyme conjugate to each well. Incubate for 10 9758 Reveal Bulk Media – 20 Hour – Contains 500 grams for
60 tests
minutes at room temperature.
11. Repeat step #9. 9760 Reveal Media – 8 Hour – Contains 20 media bottles for
20 tests
12. Wash each well 1 time with distilled water and dry by tapping
9765 Reveal Bulk Media – 8 Hour – Contains 500 grams for
out wells on a lint free towel.
56 tests
13. Add 100 µL of substrate to each well.
9770 Reveal Media – 20 Hour – Contains 20 media bottles for
14. Incubate wells for 10 minutes at room temperature.
20 tests
15. Add 100 µL of stop solution to each well.
9779 Reveal Bulk Media – Modified 8 Hour – Contains 624
16. Read optical density at 450 nm and record results.
grams for 56 tests
See package insert for complete instructions To use media other than those listed, see our Acumedia product line.

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