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Index / Bioquant / Methadone Direct / Product Detail : BQ 095D Methadone Direct
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  Price : 401   EUR
455   USD
311   GBP
1685   Zloty
53677   JPY
3093   NOK
3315   SEK
453   CHF

Product name : Methadone Direct

Catalog number : BQ 095D

Quantity: 96/kit

Availability: Yes

Supplier name : Bioquant

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More Details about


METHADONE DIRECT ELISA KIT
Catalog No. BQ 095D (96 tests)


INTENED TO USE
The METHADONE Direct ELISA Kit provides only a preliminary analytical test result. A more specific alternate chemical method must be used in order to obtain a confirmed analytical result. Gas chromatography/ mass spectrometry (GS-MS) is the preferred confirmatory method. Professional judgement should be applied to any drug of abuse test result, particularly when preliminary positive results are used.


SUMMARY AND EXPLANATION
The METHADONE Direct ELISA Kit is a specific and sensitive in-vitro test to detect the presence of Methadone and in samples such as whole blood, oral fluids, serum and urine.


PRINCIPLE OF THE TEST
The Methadone Direct ELISA Kit is based upon the competitive binding to antibody of enzyme labeled antigen and unlabeled antigen, in proportion to their concentration in the reaction mixture. A 10 ěl. aliquot of a diluted unknown specimen is incubated with a 100 ěl. dilution of enzyme (Horseradish peroxidase) labeled Methadone derivative in micro-plate wells, coated with fixed amounts of oriented high affinity purified polyclonal antibody. The wells are washed thoroughly and a chromogenic substrate added. The color produced is stopped using a dilute acid stop solution and the wells read at 450 nm. The intensity of the color developed is inversely proportional to the concentration of drug in the sample. The technique is sensitive to 25 ng/ml. The METHADONE Direct ELISA Kit avoids extraction of urine or blood sample for measurement. It employs a Methadone directed antiserum. Due to the proprietary method of orienting the antibody on the polystyrene micro-plate much higher sensitivity is achieved compared to passive adsorption. This allows an extremely small sample size, reducing matrix effects and interference with binding proteins(s) or other macromolecules.



MATERIALS NOT PROVIDED
1. Distilled or deionized water
2. Precision pipettes
3. Disposable pipette tips
4. ELISA reader capable of reading absorbance at 450nm
5. Absorbance paper or paper towel
6. Graph paper


STORAGE AND STABILITY
1. Store the kit at 2 - 8° C.
2. Keep microwells sealed in a dry bag with desiccants.
3. The reagents are stable until expiration of the kit.
4. Do not expose test reagents to heat, sun, or strong light.


WARNINGS AND PRECAUTIONS
1. Potential biohazardous materials:
The calibrator and controls contain human source components which have been tested and found non-reactive for hepatitis B surface antigen as well as HIV antibody with FDA licensed reagents. However, there is no test method that can offer complete assurance that HIV, Hepatitis B virus or other infectious agents are absent. These reagents should be handled at the Biosafety Level 2, as recommended in the Centers for Disease Control/National Institutes
of Health manual, "Biosafety in Microbiological and Biomedical Laboratories" 1984.
2. This test kit is designed for Research Use Only.
3. Do not pipette by mouth. Do not smoke, eat, or drink in the areas in which specimens or kit reagents are handled.
4. The components in this kit are intended for use as an integral unit. The components of different lots should not be mixed.
5. It is recommended that standards, control and serum samples be run in duplicate.
6. Optimal results will be obtained by strict adherence to this protocol. Accurate and precise pipetting, as well as following the exact time and temperature requirements prescribed are essential. Any deviation from this may yield invalid data.


STORAGE AND STABILTY
The expiration date of the kit is stated on the label. The kit can be expected to perform satisfactorily until the expiration date if stored in the refrigerator at 2 – 40 C.


SPECIMEN COLLECTION HANDLING
1. The Methadone specific Direct ELISA Kit is to be used with human samples, such as whole blood, oral fluids, serum, plasma and urine. has not tested all possible applications of this assay.
2. Specimens to which sodium azide has been added affect the assay.
3. Urine samples should be stored at 2 - 40C until use. Samples should be well mixed before assay. Repeated freezing and thawing should be avoided. Urine samples should be shipped refrigerated with Blue Ice or equivalent.


ASSAY PROCEDURE
All reagents must be brought to room temperature (18-260C) before use. The procedure as described below may be followed in sequence using manual pipettes. Alternatively all reagents may be added using an automated pipettor . BLOOD: For a blood cutoff of 100 ng/ml - 200 ng/ml dilute all controls and samples 1:10 in Phosphate Buffer Saline and use 10 ul of the diluted sample in each well. ORAL FLUIDS/ SALIVA: For a saliva cutoff of 25 ng/ml - 50 ng/ml dilute all controls and samples 1:10 in Phosphate Buffer Saline and use 25 ul of the diluted sample in each well. URINE: For a urine cutoff of 300 ng/ml dilute all controls and samples 1:40 in Phosphate Buffer Saline and use 10 ěl of the diluted sample in each well.
1. Dilute specimens, to the necessary range with Phosphate Buffer Saline pH 7.0. The dilution factor and volume added can be adjusted based on the laboratory’s cutoff.
2. Add 10 ěl. of appropriately diluted calibrators and standards to each well in duplicate.
3. Add 10 ěl. of the diluted specimens in duplicate (recommended) to each well.
4. Add 100 ěl. of the Enzyme Conjugate to each well. Tap the sides of the plate holder to ensure proper mixing.
5. Incubate for 60 minutes at room temperature (18-260 C) preferably in the dark, after addition of enzyme conjugate to the last well.
6. Wash the wells 6 times with 350 ěl. distilled water using either a suitable plate washer or wash bottle taking care not to cross contaminate wells. If testing samples containing abnormally high amounts of hemoglobin (some Postmortem samples), use 10 mM Phosphate buffered saline pH 7.0-7.4. This will lower potential nonspecific binding of hemoglobin to the well, thus lowering background color.
7. Invert wells and vigorously slap dry on absorbent paper to ensure all residual moisture is removed. This step is critical to ensure that residual enzyme conjugate, does not skew results. If using an automated system, ensure that the final aspiration on the wash cycle aspirates from either side of the well.
8. Add 100 ěl. of Substrate reagent to each well and tap sides of plate holder to ensure proper mixing.
9. Incubate for 30 minutes at room temperature, preferably in the dark.
10. Add 100 ěl. of Stop Solution to each well, to change the blue color to yellow.
11. Measure the absorbance at a dual wavelength of 450 nm and 650 nm.
12. Wells should be read within 1 hour of yellow color development.
The following data represent a typical dose/response curve.


The dose/response curve shown above should not be used in assay calculations. It is recommended that at least one in-house positive quality control sample be included with every assay run. A dose response curve or a cutoff calibrator should be run with every plate.

RESULTS
If the average sample absorbance is equal to or less than the average absorbance of the laboratory positive reference standard the sample is POSITIVE for Methadone. If the average sample absorbance is greater than the average absorbance of the laboratory positive reference standard the sample is called NEGATIVE for Methadone .Alternatively a dose response curve can be established by plotting standard concentration (abscissa) against corresponding absorbance (ordinate). Values for unknown samples are obtained by interpolation from the curve.

SPECIFIC PERFORMANCE CHARACTERISTICS
1. Accuracy
50 known negative urines were screened using a cutoff of 300 ng/ml of Methadone with the Methadone Direct ELISA kit. All 50 samples screened negative. 25 urine samples containing amounts of Methadone greater than 300 ng/ml, confirmed by GC/MS were screened using a cutoff of 300 ng/ml of Methadone with the Direct ELISA Kit. All 25 samples screened positive at the 300 ng/ml cutoff
2. Precision
The precision of the Methadone Direct ELISA Kit has been verified by assessment of the mean, standard deviation (SD) and coefficients of variation (CV) in data resulting from repetitive assays.
3. Intra-assay Precision
Intra-assay precision was determined with reference controls.
A 0, 125, 250 and 500 pg/well standard was assayed eight times in the same assay.


4. Specificity
The specificity of the Methadone Direct ELISA for was determined by generating inhibition curves for each of the compounds listed below The antisera cross-reactivities are listed.


5. Cross-Reactivity with Unrelated Drugs
Aliquots of a human urine matrix were spiked with the following compounds at a concentration of 50000 ng/ml. None of these compounds gave values in the assay that were equal to or greater than the assay sensitivity level (<5 ng/ml).
Acetaminophen, Acetylsalicylic acid, Amphetamine, Aminopyrine, Ampicillin, Amobarbital, Ascorbic acid, Atropine, Barbital, Benzoylecgonine, Butabarbital, Caffeine , Carbamazepine , Cocaine, Codeine , Chloroquine , Carbromal ,Desipramine, Dextromethorphan, Dextropropoxyphene, 5,5- Diphenylhydantoin, 10-11-Dihydrocarbamazepine, Diazepam, Ethosuximide , Estriol, Estrone, Estradiol, Ethotoin, Glutethimide, Hexobarbital , Ibuprofen, Imipramine, Lidocaine, LSD, Methaqualone, Methamphetamine, Metharbital , Mephenytoin, a-Methyl-a-propylsuccinimide , Mephobarbital, Methyl PEMA, Methsuximide , 4-Methylprimidone, Morphine , Meperidine, Niacinamide, Norethindrone, N-Normethsuximide, Phenobarbital , Phensuximide, PEMA, Primidone, Phencyclidine, Pentobarbital, Phenylpropanolamine, Procaine, Quinine, Secobarbital, Tetracycline, Tetrahydrozoline, THCCOOH.

Warning
All of Bio-Quant ELISA Kits have not been tested for clinical use and are not approved in the United States by the FDA for diagnostic clinical use. They are components or reagents made solely for research use, further manufacturing and export use. It is the commitment of Bio-Quant customers to receive its products solely for the purpose of exportation or research, and not for the purposes of clinical diagnostic use.
BIO-QUANT DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO THESE PRODUCTS. IN PARTICULAR Bio-Quant, INC. DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NON-INFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE OF ANY PRODUCT.

 

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