Instruction for use.
A solid-phase enzyme immunoassay for the quantitative determination of pork in human food
1.INTENDED USE
A solid-phase enzyme immunoassay for the quantitative determination of pork in food.
This kit is designed for measurement of pork in food. For possibility of use with other sample types, please, refer to Application Notes (on request). The kit contains reagents sufficient for 96 determinations and allows to analyze 43 unknown samples in duplicates.
2.SUMMARY AND EXPLANATION
Consumption and handling of pork meat is strictly prohibited by certain religious and/or legal prescriptions. Pork meat may also cause allergy and food intolerance. PORK EIA test is based on immunometric determination of porcine specific proteins by highly specific monoclonal antibodies. The test is highly sensitive and detects trace amounts of pork meat. Sensitivity does not depend on culinary processing - it is roughly the same for fresh pork meat and for prepared pork meat, including boiling, frying, stewing, smoking.
3.PRINCIPLE OF THE TEST
This test is based on two-site sandwich enzyme immunoassay principle. Tested specimen is placed into the microwells coated by specific anti-pork-antibodies. Antigens from the specimen binds to the antibodies fixed on the microwell surface. Unbound material is removed by washing procedure. Second antibodies directed towards another epitope of pork and labeled with peroxidase enzyme, are then added into the microwells. After subsequent washing procedure, the remaining enzymatic activity bound to the microwell surface is detected and quantified by addition of chromogen-substrate mixture, stop solution and photometry at 450 nm. Optical density in the microwell is directly related to the quantity of the measured analyte in the specimen.
4. WARNINGS AND PRECAUTIONS
4.1. For professional use only.
4.2. This kit is intended for in vitro diagnostic use only.
4.3. Avoid contact with stop solution containing 5,0% H2SO4. It may cause skin irritation and burns.
4.4. Wear disposable latex gloves when handling specimens and reagents. Microbial contamination of reagents may give false results.
4.5. Do not use the kit beyond the expiration date.
4.6. All indicated volumes have to be performed according to the protocol. Optimal test results are only obtained when using calibrated pipettes and microplate readers.
4.7. Do not smoke, eat, drink or apply cosmetics in areas where specimens or kit reagents are handled.
4.8. Chemicals and prepared or used reagents have to be treated as hazardous waste according to the national biohazard safety guidelines or regulations.
4.9. Do not mix reagents from different lots.
4.10. Replace caps on reagents immediately. Do not swap caps.
4.11. Do not pipette reagents by mouth.
4.12. Specimens must not contain any AZIDE compounds – they inhibit activity of peroxidase.
4.13. Safety Data Sheet for this product is available upon request directly from XEMA Co., Ltd.
4.14. The Safety Data Sheet fit the requirements of EU Guideline 91/155 EC.
5.KIT COMPONENTS
5.1. Contents of the Kit
|
Symbol
|
Description
|
Qty
|
Units
|
Colour code
|
Stability of opened/diluted components
|
1
|
SORB MTP
|
PORK EIA strips, 8х12 wells
|
polystyrene microwells coated with specific anti-pork antibodies
|
1
|
pcs
|
|
until exp.date
|
2
|
CAL 1 - 5
|
Calibrator set, 0.8 ml each. The set contains 5 calibrators: 0; 10; 30; 100; 300 U/ml
|
pork diluted in TRIS buffered preservative - 0,01% Bronidox L, 0,01% 2-Methyl-4-isothiazolin-3-one-hydrochloride; also contains red dye
|
5
|
pcs
|
red (C1 - colourless)
|
2 months
|
3
|
CONJ HRP
|
Conjugate, 11 ml
|
aqueous solution of antibodies to another epitope of pork coupled with horseradish peroxidase diluted on phosphate buffered solution with casein from bovine milk and detergent (Tween-20), contains 0,1% phenol as preservative and red dye
|
1
|
pcs
|
red
|
until exp.date
|
4
|
DIL
|
EIA buffer 11 ml
|
phosphate buffered saline with casein from bovine milk and detergent (Tween-20), contains 0,1% phenol as preservative; contains blue dye
|
1
|
pcs
|
blue
|
until exp.date
|
5
|
SUBS TMB
|
Substrate solution, 11 ml
|
ready-to-use single-component tetramethylbenzidine (TMB) solution.
|
1
|
pcs
|
colourless
|
until exp.date
|
6
|
BUF WASH 21X
|
Washing solution concentrate 21x, 22 ml
|
aqueous solution of sodium chloride and detergent (Tween 20), contains proClin300 as a preservative
|
1
|
pcs
|
colourless
|
Concentrate - until exp.date
Diluted washing solution - 1 month at 2-8 °C or 5 days at RT
|
7
|
STOP
|
Stop solution, 11 ml
|
5,0% vol/vol solution of sulphuric acid
|
1
|
pcs
|
colourless
|
until exp.date
|
8
|
N003
|
Plate sealing tape
|
|
2
|
pcs
|
|
N/A
|
9
|
K366I
|
Instruction PORK EIA
|
|
1
|
pcs
|
|
N/A
|
10
|
K366Q
|
QC data sheet PORK EIA
|
|
1
|
pcs
|
|
N/A
|
Equipment and material required but not provided
- Distilled or deionized water;
- Automatic or semiautomatic multichannel micropipettes, 100-250 µl, is useful but not essential;
- Calibrated micropipettes with variable volume, range volume 25-250 µl;
- Calibrated microplate photometer with 450 nm wavelength and OD measuring range 0-3.0.
- Preservative for samples (XEMA Cat. # S075Z) – optional
- Balance with precision of 0.1 g (for weighting meat products)
5.3. Storage and stability of the Kit
Store the whole kit at 2 to 8 °C upon receipt until the expiration date.
After opening the pouch keep unused microtiter wells TIGHTLY SEALED BY ADHESIVE TAPE (INCLUDED) to minimize exposure to moisture.
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