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EGT Test Kit
EGT Test Kit
This page contains information on EGT Test Kit for veterinary use.
The information provided typically includes the following:
EGT Test Kit Indications
Warnings and cautions for EGT Test Kit
Direction and dosage information for EGT Test Kit
Egt Test Kit
This treatment applies to the following species:
Cats
Dogs
Manufacturer: PRN Pharmacal
Description
A complete test kit to detect the presence of ethylene glycol in canine or feline blood, plasma or serum.
Components
Tubes: Each test pack contains several tubes with colored caps. It may be advisable for the first-time user to mark the tubes to correspond with cap color to avoid any possible confusion.
Pipets: Several calibrated (1 mL with 0.25 mL divisions) plastic pipets are included in each test pack that are to be used for transfer operations. Dispose of all pipets when instructed to do so in order to avoid cross-contamination.
Materials required, but not included: Distilled water, centrifuge tubes, centrifuge.
EGT Test Kit Indications
Peak blood levels of ethylene glycol are reached in one to four hours post ingestion at which time the early clinical signs of intoxication are apparent (depression, ataxia especially in the rear, polydipsia, polyuria, metabolic acidosis, and serum hyperosmolality). Dogs exhibiting any of these signs of toxicosis should be tested immediately. Additionally, dogs presented up to twelve hours post ingestion may be tested. Beyond twelve hours post ingestion test results will be marginal because most of the ethylene glycol will have been excreted or metabolized. Diagnosis should then be made on the basis of history, metabolic acidosis, and the presence of calcium oxalate and especially hippuric acid crystals in the urine. The determinations of the anion and osmolal gaps as well as serum glycolic acid level are also useful in belated diagnosis of antifreeze poisoning. Unfortunately, these tests often are not readily available.
Test Principles
Blood cells are lysed in water and blood proteins are denatured and precipitated by the addition of sodium tungstate and dilute sulfuric acid. Centrifugation removes precipitated blood components leaving ethylene glycol and other small molecules in solution. Ethylene glycol is oxidized by periodate to formaldehyde which forms a colored complex with 4-Amino-3-hydrazino-5-mercapto-1,2,4-triazole at alkaline pH.3
Test Procedure
1) Draw blood using EDTA or heparin as an anticoagulant. One-half mL of whole blood is required for the test. Serum or plasma may be used if more convenient. (See caution below!)
2) Add 3 mL deionized or distilled water to a centrifuge tube that can hold a total of 5 mL. (If a 5 mL centrifuge tube is not available, use a convenient container such as a small paper cup for steps 2-5 and then distribute contents to two or more centrifuge tubes of any size.)
3) Using one of the plastic transfer pipets provided, add exactly 0.5 mL of whole blood, serum, or plasma to the water and mix completely. Reserve the pipet for use in steps 4 and 5.
4) Carefully open the tube with the white cap, and add the entire contents to the blood-water mixture. Use the plastic pipet saved from step 3 to mix completely.
5) Carefully open the tube with the lavender cap, and add the entire contents to the blood-water mixture. Again use the plastic pipet to mix completely and all the mixture to stand for 5 minutes before proceeding. Dispose of the pipet properly now.
6) Distribute the test sample to appropriate centrifuge tubes (not provided), and spin at blood speed for 5 minutes. A brownish-red pellet of precipitated blood proteins with a water-clear, colorless supernatant should result. Disregard any slight amount of material floating on top of the supernatant. Repeat the test using serum or plasma if a colorless supernatant is not obtained with whole blood.
7) Use a new, clean plastic pipet to transfer exactly 1 mL of the clear test sample supernatant to the yellow-capped reaction tube provided. Avoid any floating material. Dispose of the pipet properly now.
8) Use a new, clean plastic pipet to transfer exactly 0.25 mL of the oxidizer solution (blue-capped tube) to the test sample (yellow-capped tube). Do not allow the pipet to contact the test sample solution. Use the same pipet to transfer exactly 0.25 mL of the oxidizer solution (blue-capped tube) to the control tube (green-capped tube). Securely recap the reaction tube and control tube, shake vigorously, and allow the oxidation reaction to proceed for 10 minutes. Dispose of the pipet properly now.
Important: Be sure that you have added oxidizer solution to both the reaction and control tube.
9) The color reagent is supplied in a pair of red-capped tubes. One tube contains a diluent, the other contains a powder capsule. Activate the color reagent by using a plastic pipet to carefully combine the liquid with the powder. Reserve the pipet for use in step 10.
10) Use the pipet saved from step 9 to transfer 2 mL of the color reagent (red-capped tube mixture) to the yellow-capped reaction tube. Do not allow the pipet to contact the test sample solution. Also transfer 2 mL of the color reagent to the green-capped control tube. Make sure both tubes are recapped securely and shake vigorously. Observe results in 5 minutes. Dispose of the pipet properly now.
Test Interpretation
(See Caution section below!)
A definite reddish-violet (lavender) color should develop in the green-capped control tube within 5 minutes. Color development in the yellow-capped reaction tubs as intense or more so than the control tube is indicative of blood ethylene glycol levels of greater than 0.5 ppt (the standard is 50 µg/mL) and antifreeze poisoning. The color of a strongly positive sample may be very dark. Very faint color may develop in the reaction tube with normal blood due to slight interference from blood aldehydes, but this color should be much less than that of the positive control tube. Commonly, a normal sample will yield a faint yellowish color which may fade to colorless or extremely faint reddish-violet after standing for 30 minutes to an hour.
Please read the caution below concerning false positive reactions due to propylene glycol and glycerol. A false positive test will also result from metaldehyde, a component in some slug baits. If color of a different hue than that of the control tube develops, precipitates form, or no color develops in the control tube, disregard results and contact the manufacturer. If problems should arise, please include the lot number of your test kit in any correspondence.
EGT Test Kit Caution(s)
Draw blood for this test before administration of any preparations containing propylene glycol or glycerol. Failure may result in a false positive test reaction. (Note: Some activated charcoal suspensions and semi-moist diets contain propylene glycol. Propylene glycol may not be listed in ingredients; if in doubt, contact the manufacturer.
Some of the materials in the kit are caustic. No solutions should be allowed to come in contact with the skin or eyes. If contact should occur, flush with copious amounts of water. Use the pipets provided for all operations--do not mouth pipet.
Discussion
A general understanding of the metabolism of ethylene glycol is crucial for accurate diagnosis of poisoning and for choosing a therapy regimen. The metabolic degradation of ethylene glycol takes place in the liver. Alcohol dehydrogenase converts ethylene glycol slowly to glycoaldehyde which is then quickly converted to glycolic acid. Glycolic acid concentration builds because all metabolic pathways that can degrade this compound are slow acting.
Most of the harmful effects of ethylene glycol poisoning are associated with glycolic acid (i.e. severe metabolic acidosis) and metabolites of glycolic acid (i.e. calcium oxalate crystals). Clinical signs and mortality correlate with urinary glycolate concentration. As the ethylene glycol is metabolized, animals often appear to recover from the direct effects of the poisoning only to rapidly deteriorate as glycolic acid concentration builds. Grauer et al.2 followed the course of toxicosis in fifteen dogs given 9.5 mL of ethylene glycol/kg body weight. Depression, ataxia especially in the rear, polydipsia, polyuria, metabolic acidosis, and serum hyperosmolality were found at one to three hours, whereas calcium oxalate crystalluria was first observed at six hours, and renal function was not obviously impaired until 48 hours.
Most treatment regimens are based on some combination of ethanol, bicarbonate, and fluids.1 Ethanol competitively inhibits alcohol dehydrogenase thus reducing the metabolism of ethylene glycol into more toxic compounds. [Note: Recently, 4-methylpyrazole has been recommended as an inhibitor of alcohol dehydrogenase that does not cause the undesirable CNS effects of alcohol.4] Correction of metabolic acidosis (blood pH < 7.3) is accomplished with sodium bicarbonate in fluid therapy to maintain urine pH at approximately 7.5 to 8.0. Ethanol is not indicated when animals are presented at more than 18 hours because most of the ethylene glycol would already be metabolized by alcohol dehydrogenase and a competitive block of this enzyme would be of no use. Supportive fluid and bicarbonate therapy is appropriate at this time. It should be understood that ethanol treatment will cause blood levels of ethylene glycol to remain elevated for prolonged times (because metabolism by alcohol dehydrogenase is blocked), and treatment periods must be lengthy (up to 72 hours) to allow time for excretion of the ethylene glycol.
Clearly, rapid diagnosis and minimal delay in treatment are positive factors in the prognosis of antifreeze poisoning. The PRN Ethylene Glycol Test Kit is useful in determining toxicosis one to twelve hours after ingestion. An informed diagnosis can be made quickly and the appropriate treatment can be initiated without delay because the test results can be read within 30 minutes. Sequential tests may be made during the course of ethanol treatment to follow blood levels of ethylene glycol.
References
Available upon request.
Presentation
4 tests per kit.
Nac No.
10900030
PRN PHARMACAL
8809 ELY ROAD, PENSACOLA, FL, 32514
Telephone:
850-476-9462
Order Desk:
800-874-9764
Fax:
850-476-7087
Website:
www.prnpharmacal.com
Email:
gcushing@prnpharmacal.com
Every effort has been made to ensure the accuracy of the EGT Test Kit information published above. However, it remains the responsibility of the readers to familiarize themselves with the product information contained on the EGT Test Kit product label or package insert.
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