Procedures & Specimen Requirements

Selected Test Procedures

ACTH Stimulation: Canine/Feline
Test Code: 126
Availability: Daily
Submission Requirements: 0.5ml serum per specimen (do not submit EDTA plasma)
Suggested Protocol: Fast the cat/dog approximately 12 hrs (preferred, not essential) and collect 3 ml of blood. Centrifuge and separate at least 0.5ml of serum for a resting cortisol.
Canine: Synthetic ACTH:
1) Administer one vial (0.25mg) of synthetic ACTH IM, regardless of the dog’s size. Alternatively, administer 5 ug/kg of synthetic ACTH IM.
2) Collect 0.5ml of serum one hour after injection for a post-cortisol.
Feline: Synthetic ACTH:
1) Administer 0.125mg( half-vial) of synthetic ACTH IM.
2) Collect 0.5ml of serum 30 minutes and 60 minutes after injection for post cortisols.
Label the tubes with the patient’s last name and either “Pre” or “Post 2 hrs”

Aspergillus Antibody AGID (non-avian)
Test Code: 500
Availability: Send out: 3-5 days.
Submission Requirements: 1 ml serum
Storage: Refrigerate
Interferences: Marked hemolysis, lipemia
Interpretation: Positive or Negative

Baermann Test (lungworms)
Test Code: 808
Availability: 2-3 days
Submission Requirements: 2-5 g fresh feces in sterile container; do not use formalin
Storage: Refrigerate
Interpretation: No parasites seen or parasites identified
Comment: Do not allow specimen to touch ground or be exposed to flies.

Bile Acids Panel (Pre- and Postprandial)
Test Code: 119
Availability: Daily
Submission Requirements: 0.5 ml serum per sample time.
Storage: Refrigerate
Suggested Protocol:
1) Fast the dog or cat approximately 12 hours and obtain a fasting (preprandial) serum specimen of at least 1ml. Label the tube Fasting or Preprandial.
2) Ideally, feed the animal a high-fat meal in order to stimulate gallbladder contractions. Minimum amount of food is 2 teaspooons for small patients (<10lb) and 2 tablespoons for large patients.
3) Two hours after feeding, obtain a postprandial serum specimen of at least 1ml, and label the tube Postprandial.
Interferences: Hemolysis, lipemia, and icterus
Canine and Feline: Fasting or preprandial bile acid concentrations >10 umol/L and postprandial bile acids concentrations >20 umol/L are associated with hepatobiliary disease.

Blood Typing: Canine/Feline
Test Code: Canine: 310/ Feline 311
Availability: Daily
Submission Requirements: 1ml whole blood (EDTA)
Storage: Refrigerate
Interpretation: Positive or Negative (DEA 1.1, neg and DEA 1.1 pos. in dogs A, B, or AB in cats)

Bromide (KBR)
Test Code: 378
Availability: Results available daily.
Synonym: Potassium bromide, KBR, sodium bromide
Submission Requirements: 1ml serum (WTT); do not submit in SST
Storage: Refrigerate
Interferences: Hemolysis, icterus, and SST gel
Therapeutic Monitoring:
• Nonloading dose treatment protocol: measure bromide concentration one month into treatment and again at 2 to 3 months (steady-state concentration). 1 month bromide concentration approximates ½ of future steady state concentrations; therefore, dose may be adjusted proactively.
• Loading dose treatment protocol: measure bromide concentration immediately post-loading dose, 1 month into treatment, and again in 2-3 months. If post-loading and one-month concentrations vary more than 10% from each other, maintenance dose can be modified accordingly.
• Recommend verification that bromide concentrations are within therapeutic range before decreasing Phenobarbital dose.
Therapeutic Range:
Canine: 1-2.mg/mL (100-200 mg/dl) in combination with Phenobarbital or 2.5-3.0 mg/mL (250-300mg/dl) as sole anticonvulsant (without Phenobarbital)
Steady-state drug concentrations are expected in 3-4 months. Due to patient variability, therapeutic drug concentrations should be interpreted in combination with patient’s clinical signs (See also Phenobarbital).

CALP
Test Code: 316
CALP/ALP
Test Code: 431
Availability: Daily
Submission Requirements: 0.5ml serum
Storage: Refrigerate
Interferences: Avoid hemolysis/lipemia (Moderate to marked hemolysis will invalidate assay)
Interpretation: (Provided by Dr. Walter Hoffman)
The corticosteroid induced isoenzyme of alkaline phosphate (CALP) is a unique enzyme found only in dogs. Normal dogs have little or none of this enzyme. Its presence in serum is often observed in dogs with Cushing’s disease, iatrogenic Cushing’s syndrome and in some older dogs with chronic disease. In dogs with greater than 30 U/L CALP activities, an examination for clinical signs of Cushing’s disease as well as an investigation of history of prior treatment with glucocorticoids should be made. Additional diagnostic testing for Cushing’s disease may be warranted. The sensitivity for CALP in the diagnostic of Cushing’s disease is nearly 100% but the specificity is in the order of 85%. In other words, Cushing’s disease is very unlikely if the CALP activity is not increased. However, there are false positives (especially in older dogs with chronic disease) so that increased CALP activity cannot be used as the sole means for diagnosis of Cushing’s disease unless very clearly supported with clinical signs.

Coombs: Canine/Feline
Test Code: 324
Availability: Daily
Submission Requirements: 1ml whole blood (EDTA)
Storage: Refrigerate
Interferences: Steroid therapy, severe auto-agglutination
Interpretation: Positive or negative with degree of agglutination.

Cryptococcus antigen
Test Code: 501
Availability: Results available in 2-5 days
Submission Requirements: 1 ml serum or CSF (SST); serum in SST preferred. *Do not submit plasma. Do not use plastic tubes.
Storage: Refrigerate
Interferences: RBC’s, marked hemolysis, lipemia turbidity, microbial contamination.
Interpretation: Screen performed on undiluted serum; positives titered to endpoint.
Latex gloves do not cause interference with this test.

Dexamethasone Suppression, Canine High-dose (Baseline, 4-hours, 8-hours)
Test Code: 300
Availability: Daily
Method: Chemiluminescence
Submission Requirements: 0.5mL serum (SST) per specimen
Storage: Refrigerate. Freeze if stored more than 3 days.
Information: Sampling time: Baseline, 4-hour, and 8-hour post
Drug: Use either dexamethasone sodium phosphate or dexamethasone in polyethylene glycol.
Dosage: 0.1mg/kg IV
Interpretation:
Baseline cortisol: 1.0-5.0 ug/dL
If the 4-hour and/or 8-hour post cortisols are <50% of the baseline, the diagnosis is pituitary dependent hyperadrenocorticism. (80% of pituitary dependent hyperdrenocorticism cases have 4-hour or 8-hour cortisols < 50% of the baseline.) If the 4-hour and/or 8-hour post cortisols are >50% of the baseline the diagnosis is either adrenal dependent or pituitary dependent hyperadrenocorticism. This assay is used for previously diagnosed canines and is used to distinguish pituitary from adrenal-dependent hyperadrenocorticism.

Dexamethasone Suppression, Canine Low-dose (Baseline, 4-hours, 8-hours)
Test Code: 127
Availability: Daily
Method: Chemiluminescence
Submission Requirements: 1ml serum each (SST)
Storage: Refrigerate. Freeze if stored more than 3 days.
Information: Sampling time: Baseline, 4-hour, and 8-hour post. Drug: Use either dexamethasone sodium phosphate or dexamethasone in polyethylene glycol.
Dosage: 0.01mg/kg IV
Interpretation:
• Baseline cortisol: 1.0-5.0 ug/dL
• 4-hour and 8-hour post cortisol: <1.4 ug/dL
• Canine: The 8-hour post sample is used to screen for hyperadrenocorticism: 8-hour post cortisol >1.4 ug/dL supports hyperadrenocorticism. The 4-hour post sample is used to help differentiate adrenal dependent versus pituitary dependent hyperadrenocorticism: 4-hour cortisol <1ug/dL supports pituitary dependent hyperadrenocorticism; if the 4-hour cortisol is >1 ug/dL, then additional tests are needed to differentiate adrenal dependent and pituitary dependent hyperadrenocorticism.
• Feline: Dexamethasone suppression assays are not as easily interpreted in the cat as in the dog. Cortisol values of >1.5 ug/dL in either of the two post dexamethasone samples suggest hyperadrenocorticism.

Distemper and Parvo Titer (canine)
Test Code: 8661
Availability: Daily
Includes: Distemper and Parvovirus antibody
Submission Requirements: 1mL serum (SST)
Storage: Refridgerate up to 4 days.
Interpretation: A positive distemper ELISA result correlates with an SN antibody titer > 1:16, indicating probable protective immunity. A positive parvovirus ELISA result correlates with an IFA antibody titer >1:80, indicating probably protective immunity. A negative result indicates a low antibody level, and a booster vaccination should be considered.

Ethylene Glycol (qualitative)
Test Code: 345
Availability: Daily
Submission Requirements: 2 ml whole blood (EDTA)
Storage: Refrigerate
Interpretation: Positive or Negative
This test is most useful when specimen is drawn within 12 hours of ingestion, and before treatment.

Fecal Occult Blood
Test Code: 347
Availability: Daily
Method: Guaiac test
Submission Requirements: 1 tablespoon fresh feces
Storage: Refrigerate
Suggested Protocol: Avoid red meat, (liver, beef, lamb) and their by-products for 2-3 days. While remaining on this special diet, collect a stool sample once a day for 3 days. Pool the samples together and send it in on third day.
Interferences: Aspirin, corticosteroids, iron supplements, dietary blood, horseradish, red meat diet.
Interpretation: Positive or Negative.

FeLV Antigen IFA
Test Code: 351
Availability: Results available in 7-10 days
Synonym: Feline leukemia virus
Submission Requirements: 2 air-dried UNSTAINED bone marrow or peripheral blood smears or 1 mL whole blood (EDTA)
Storage: Refrigerate up to 4 days, store slides at room temperature.
Interferences: Decreased platelets, decreased WBC count, poor slide quality, eosinophilia, slight hemolysis.
Interpretation: Positive or negative. Vaccination does NOT interfere with this test.

Fibrogen Estimate by heat precipiatation
Test Code: 352
Availability: Daily
Submission Requirements: 1ml whole blood (LTT)
Storage: 24 hours at 2°-8° C.

Fructosamine
Test Code: 354
Availability: Daily
Submission Requirements: 1ml serum or plasma (SST or GTT); nonhemolyzed, fasted sample.
Storage: Refrigerate for up to 4 days
Interference: Moderate to marked hemolysis, icterus
Interpretation: This test reflects glycemic status over the previous 1-3 weeks. It may be helpful in differentiating elevated glucose values due to stress versus diabetes mellitus and in monitoring therapy for diabetes mellitus.

Fungal Culture
Test Code: 334
Availability: Test may require 20-30 day incubation.
Submission Requirement: Tissue, hair or body fluid in a sterile container (WTT) preferred. Culturette from ear acceptable. Do not use formalin.
Storage: Refrigerate for up to 4 days
Interpretation: Results reported as no fungal growth or fungal identification. Cultures evaluated as needed and finalized within four weeks.

Fungal Serology Panel
Test Code: 121
Availability: Results available in 5-7 days.
Method: AGID
Test includes: Blastomyces, Histoplasma, Coccidiodes and Aspergillus antibodies.
Submission Requirements: 3 ml serum (SST)
Storage: Refrigerate for up to 4 days
Interferences: Anticoagulants (EDTA or citrate) or preservatives
Interpretation: Positive or negative; endpoint titer for Coccidiodes-positive. Test requires 48 hour incubation.

Iron & Iron Total Binding Capacity, % Saturation
Test Code: 503
Availability: Results available in 4-7 days
Test Includes: Serum iron, total iron-binding capacity (TIBC), % saturation
Submission Requirements: 2ml serum (SST); Centrifuge within two hours of blood collection, remove serum from blood tube, and then place into plastic tube (WTT) for freezing.
Storage: Frozen; Refrigerate for up to 4 days
Interferences: Hemolysis, lipemia
Interpretation: This panel is commonly used for the diagnosis of iron deficiency or anemia of inflammatory disease. Either of these syndromes can demonstrate serum iron levels that range from normal to low. In cases of iron deficiency, TIBC will often be high to normal, and serum ferritin will be low. Patient’s afftected by anemia of inflammatory disease will have a normal to low TIBC and normal to high serum ferritin.

Lead Level
Test Code: 367
Availability: Results availabe in 3-5 days
Method: Atomic absorption
Submission Requirements: 1 mL whole blood (EDTA) or heparinized whole blood (GTT) *DO NOT submit serum for this test.
Storage: Refrigerate
Interferences: Incorrect tube

Partial Thromboplastin Time (PTT)
Test Code: 383
Availability: Daily
Synonym: PTT, APTT
Submission Requirements: Citrated plasma (BTT). Tube must be filled as much as vacuum will allow. Invert several times to mix anticoagulant and blood and then centrifuge immediately. Separate plasma from cells. Transfer plasma to WTT, not another BTT. Label tube “citrated plasma” along with client’s last name.
Storage: Refrigerate for up to 4 days, indefinitely if frozen.
Interferences: Incorrect ration of citrate to whole blood, clotted specimen, noncitrate anticoagulant.

Phenobarbital
Test Code: 374
Availability: Daily
Submission Requirements: 1ml serum (WTT), do not use SST
Storage: Refrigerate for up to 4 days
Interference: Moderate to marked lipemia, icterus, SST gel
Interpretation:
• Subtherapeutic level: <15 ug/ml
• Therapeutic level: 15-40 ug/ml
• Possible toxic level: >40 ug/ml
• The biological half-life of Phenobarbital in the dog is generally 56 hours. The steady state is generally only achieved after 2-3 weeks of drug therapy. Due to patient variability (sensitivity and tolerance) serum Phenobarbital values must be evaluated in conjunction with the patient’s clinical signs.
If patient is on both potassium bromide and Phenobarbital, we recommend monitoring serum levels of both drugs.

Progesterone
Test Code: 379
Availability: Daily
Submission Requirements: 1.5 mL serum (WTT) DO NOT USE SST
Method: Chemiluminescence
Storage: Refrigerate for up to 4 days
Interference: Lipemia, EDTA
Interpretation:
(Canine): Serum progesterone in the dog should be <1ng/ml in proestrus. When progesterone rises to >2ng/ml, it is associated with the pre-ovulatory LH surge. Counting the day that this rise occurs as day 0, the fertile window occurs from day 2 to day 7.

Prothrombin Time (PT)
Test Code: 381
Availability: Daily
Submission Requirements: Citrated plasma (BTT). Tube must be filled as much as vacuum will allow. Invert several times to mix anticoagulant and blood and then centrifuge immediately. Separate plasma from cells. Transfer plasma to WTT, not another BTT. Label tube “citrated plasma” along with client’s last name.
Storage: Refrigerate for up to 24 hours, indefinitely if frozen
Interference: Incorrect ratio of citrate/whole blood. Clotted specimen, noncitrate anticoagulant.

Rheumatoid Factor
Test Code: 504
Availability: Results available in 2-3 days.
Synonym: RF, rheumatoid arthritis
Submission Requirements: 1ml serum
Storage: Refrigerate for up to 4 days
Interferences: Osteoarthritis, fibrositis, polyarteritis nodosa, EDTA
Interpretation: Positive or negative

Stone Analysis
Test Code: 394
Availability: Results available in 1-2 weeks
Submission Requirements: Calculi in a clean, dry container
Storage: Room Temperature
• Quantitative stone analysis is recommended over the qualitative analysis. Quanitiative analysis gives the percent of each mineral present.

T4, Total: Canine/Feline
Test Code: 387
Availability: Daily
Method: Chemiluminescence
Synonym: Thyroxine, tetraiodothyronine
Submission Requirements: 1 ml serum (SST)
Storage: Refrigerate for up to 4 days
Interference: Lipemia, hemolysis, anticoagulants
Interpretation: Reference values are provided as a general guideline, but each case must be individually evaluated considering numerous variables, including age, drug therapy and disease state.
A post-pill blood sample should be obtained 4-6 hours after thyroid supplement administration to monitor treatment efficacy.

Testosterone
Test Code: 818
Availability: Results available in 3-8 days
Method: RIA
Submission Requirements: 1 ml serum
Storage: Refrigerate for up to 4 days
Interferences: EDTA
Interpretation:
Canine:
Male, intact: 1-7 ng/ml
Male, neutered: <0.1 ng/ml
Female: <0.1 ng/ml
Feline:
Male, intact: 1-6 ng/ml
Male, neutered: < 0.5 ng/ml
Female: <0.2 ng/ml
Equine:
Stallion, breeding season 1-4 ng/ml
Stallion, nonbreeding season <1.0 ng/ml
Gelding <0.2ng/ml
Mare: <0.1 ng/ml

Toxoplasma (IgG and IgM)
Test Code: 391
Availability: Results available in 3-7 days
Test includes: Toxoplasmosis titer IgG and toxoplasmosis titer IgM
Submission Requirement: 1ml serum (SST)
Storage: Refrigerate for up to 4 days; freeze for longer storage.
Interferences: Hemolysis, lipemia
Interpretation: IgG titer and IgM titer given individually. Approximately 30% of dogs and cats in the US have antibodies to T. gondii. A positive titer indicates exposure, but does not confirm infection nor clinical diagnosis. Tentative diagnosis requires demonstration of a high IgM titer (>256) or a fourfold (or greater) increase in IgG titer in 2-3 weeks.
Test requires 24 hours incubation. Recommended test for canine and feline patients.

TSH, Endogenous-Canine
Test Code: 331
Availability: Daily
Storage: Refrigerate for up to 4 days
Interference: Hemolysis, anticoagulants
Interpretation: Increased canine TSH values (greater than 2.5 times normal) may occur in dogs with low T4 and untreated primary hypothyrodism. Sick euthyroid dogs are expected to have low normal TSH concentrations with low T4.

Urine Protein/Creatinine Ratio
Test Code: 396
Availability: Daily Test
Includes: Urine protein, urine creatintine, protein: creatinine ratio
Normal Range: Dogs and Cats: <1.0
Submission Requirements: 1ml urine in RTT or WTT (No additives)
Storage: Refrigerate for up to 4 days
Interferences: hematuria, pyuria

Von Willebrand Factor
Test Code: 814
Availability: Results available in 2-3 days
Submission Requirements: One plastic citrated blood tube (BTT) must be filled as much as vacuum will allow. Invert several times to mix anticoagulant and blood, then centrifuge immediately. Separate plasma from cells. Use plastic pipette to transfer plasma to plain plastic tube and freeze. Label tube “citrated plasma” along with client’s last name.
Storage: Store plasma frozen
Interferences: Hemoylsis, inclusion of RBC’s in plasma, or clotted specimen
Plasma should not touch glass because it will cause clotting factor activation.
Interferences: vWF: AG = Von Willebrand Factor antigen measured in an ELISA
70-180% = Normal range for vWF:Ag
50-69% = Borderline normal or indeterminate range for vWF:Ag
<50% = Abnormal or carrier range for vWF:Ag