| Trade - Want to Buy |
| Topic | E. coli O157:H7 ELISA Test Kit |
| Name | NICOLE |
| Company | REAGEN |
| Address | 102 Commerce Dr. Unit 8, Moorestown, NJ 08057 |
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| Country | USA |
| Date Created | 9/28/2013 1:20:27 PM |
| Message | Product Description
REAGEN™E. coli O157:H7 ELISA Test Kit is designed for routine screening for E. coli O157H7 from cultured food and feed samples. E. coli serotype O157:H7 is a gram-negative rod-shaped bacterium.The letter "O" in the name refers to the somatic antigen number, whereas the "H" refers to the flagella antigen. This is one of hundreds of serotypes of the bacterium E. coli. While most strains are harmless and normally found in the intestines of mammals, this O157:H7 strain may produce Shiga-like toxins,cause severe illness, and is a member of a class of pathogenic E. coli known as enterohemorrhagic E.coli or EHEC. People infected with enterohemorrhagic E. coli often develop hemorrhagic colitis and/or hemolytic-uremic syndrome. The sources of contamination include raw ground beef, unpasteurized milk,untreated water, juice, raw sprouts, lettuce, and salami.
The detection of E. coli O157:H7 in food samples is often laborious and time-consuming. Several days are required to culture the bacteria from a sample followed by a final isolation and identification by biochemical and serological techniques. The REAGEN™ E. coli O157:H7 ELISA Test Kit provides a simple, rapid, sensitive and cost-effective enzyme immunoassay (ELISA) screening method, which enables government agencies, seafood processors, as well as quality assurance organizations, to detect positive and negative E. coli O157:H7 samples to as low as 105 cells/mL level.
Procedure Overview
The method is based on a colorimetric ELISA assay. The monoclonal antibody against E. coli O157:H7 has been coated in the plate wells. During the analysis, the sample is added. If E. coli O157:H7 is present in the sample, it will bind with the antibody. The secondary antibody, tagged with a peroxidase enzyme, targets the E. coli O157:H7 that is complexed to the primary antibody coated on the plate wells.The resulting color intensity, after addition of substrate, is related to the amount of E. coli O157:H7 in the sample.
Kit Contents, Storage and Shelf Life
REAGEN™E.coli.0157:H7 ELISA Test Kit has the capacity for 96 determinations or testing of 42 samples in duplicate (assuming 12 wells for standards). Return any unused microwells to the foil
bag and reseal them with the desiccant provided in the original package. Store the kit at 2-8C*. The shelf life is 12 months when the kit is properly stored.
Kit Contents Amount Storage
E. coli O157:H7 antibody-coated Plate 96well plate(8wells x12 strips) 2-8C
1X E. coli O157:H7 Negative Control 1 mL -20C
1X E. coli O157:H7 Positive Control 1 mL -20C *
50X HRP-Conjugated Antibody #2 50 Lx3 -20C *
Antibody #2 Diluent **
20X Wash Solution **
Stop Buffer **
TMB Substrate ** 20 mL
30 mL
20 mL
10 mL 2-8C
Sensitivity
The kit can detect 105cells/mL.
Specificity
The kit can broadly detect E. coli O157:H7 antigen.
Required Materials Not Provided With the Kit
Microtiter plate reader (450 nm)
Incubator
Vortex mixer (e.g. Gneie Vortex mixer from VWR)
10, 20, 100 and 1000 L pipettes
Multi-channel pipette: 50-300 L (Optional)
Modified TSB Broth (mTSB): Consisting of Trypticase soy broth (TSB) supplemented with Casamino Acids (10 g/liter) and K2HPO4 (1.5 g/liter)
Warnings and Precautions
The standards contain E.coli.0157:H7 . Handle with particular care.
Do not use the kit past the expiration date.
Do not intermix reagents from different kits or lots except for components with the same part No’s within their expiration dates. ANTIBODIES AND PLATES ARE KIT- AND LOT-SPECIFIC. Make sure that the antibody #2 and diluent are mixed in correct volumes.
Try to maintain a laboratory temperature of 20°–25°C (68°–77°F). Avoid running assays under or near air vents, as this may cause excessive cooling, heating and/or evaporation. Also, do not run assays in direct sunlight, as this may cause excessive heat and
evaporation. Cold bench tops should be avoided by placing several layers of paper towel or some other insulation material under the assay plates during incubation.
Make sure you are using only distilled or deionized water since water quality is very important.
When pipetting samples or reagents into an empty microtiter plate, place the pipette tips in the lower corner of the well, making contact with the plastic.
Incubations of assay plates should be timed as precisely as possible. Be consistent when adding standards to the assay plate. Add your standards first and then your samples.
Add standards to plate only in the order from low concentration to high concentration as this will minimize the risk of compromising the standard curve.
Always refrigerate plates in sealed bags with a desiccant to maintain stability. Prevent condensation from forming on plates by allowing them equilibrate to room temperature (20 – 25C / 68 – 77F) while in the packaging.
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