Procedures & Specimen Requirements

Selected Test Procedures

Procedures and Specimen Requirements

ACTH Stimulation: Canine/Feline
Test Code: 126
Availability: Results available daily
Submission Requirements: 0.5mL serum per specimen (do not submit EDTA plasma)
Suggested Protocol: Fast the patient approximately 12 hrs (preferred, not essential) and collect 3mL of blood. Centrifuge and separate at least 0.5mL of serum for a resting cortisol.
Canine: Synthetic ACTH:
1) Administer 5µg/kg of synthetic ACTH IV. (Maximum of one vial (250 µg/0.25 mg)
2) Collect 0.5mL of serum (SST) one hour or two hours after injection for a post­cortisol.
Feline: Synthetic ACTH:
1) Administer 5µg/kg of synthetic ACTH or 0.125mg (half­vial) of synthetic ACTH IM.
2) Collect 0.5mL of serum (SST) 30 minutes and 60 minutes after injection for post cortisols.
* Label the tubes with the patient’s last name and either “Pre,” and “Post 1 hr, and/or Post 2 hr or Post 30 and 60 minutes.”

Aspergillus Antibody AGID (non­avian)
Test Code: 500
Availability: Results available in 3­5 days
Submission Requirements: 1mL serum
Storage: Refrigerate
Interferences: Marked hemolysis, lipemia
Interpretation: Positive or Negative

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

Bile Acids Panel (Pre­ and Postprandial)
Test Code: 119
Availability: Results available daily
Submission Requirements: 0.5mL serum (SST) per specimen
Storage: Refrigerate
Suggested Protocol:
1) Fast the animal 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 high­fat meal in order to stimulate gallbladder contractions. Minimum amount of food is 2 teaspooons for small patients (<10lb) and 2 tablespoons for large patients.
2) 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: Results available 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 (steady­state concentration). 1 month bromide concentration approximates ½ of future steady state concentrations; Therefore, dose may be adjusted proactively.
­ Loading dose treatment protocol: 1 month into treatment, and again in 2­3 months. If postloading 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: 1­2.mg/mL (100­200 mg/dl) in combination with Phenobarbital or 2.5 – 3.0 mg/mL (250­300mg/dl) as sole anticonvulsant (without Phenobarbital)
* Steady­state drug concentrations are expected in 3­4 months. Due to patient variability, therapeutic drug concentrations should be interpreted in combination with patient’s clinical signs. (Also see Phenobarbital)

CALP
Test Code: 316
CALP/ALP
Test Code: 431
Availability: Results available daily
Submission Requirements: 0.5mL serum (SST)
Storage: Refrigerate
Interferences: Avoid hemolysis/lipemia (Moderate to marked hemolysis will invalidate assay)
Interpretation:
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 diagnosis 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: Results available daily
Submission Requirements: 1mL whole blood (EDTA)
Storage: Whole blood is stable for 48 hours refrigerated
Interferences: Steroid therapy, severe auto­agglutination
Interpretation: Positive or Negative with degree of agglutination

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

Dexamethasone Suppression, Canine/ Feline High­dose (Baseline, 4­hours, 8­hours)
Test Code: 300
Availability: Results available daily
Method: Chemiluminescence
Submission Requirements: 0.5mL serum (SST) per specimen
Storage: Refrigerate. Freeze if stored more than 3 days
Information: Sampling time: Baseline, 4­hour, and 8­hour post
Drug: Use either dexamethasone sodium phosphate or dexamethasone in polyethylene glycol.
Dosage: 0.1mg/kg IV
Interpretation:
Baseline cortisol: 1­5ug/dL
If the 4­hour and/or 8­hour post cortisols are < 50% of the baseline, the diagnosis is pituitary dependent hyperadrenocorticism. (80% of pituitary dependent hyperdrenocorticism cases have 4­hour or 8­hour cortisols < 50% of the baseline.) If the 4­hour and/or 8­hour post cortisols are > 50% of the baseline the diagnosis is either adrenal cortical tumor dependent or pituitary dependent hyperadrenocorticism. This assay is used for previously diagnosed canines and is used to distinguish pituitary from adrenal cortical tumor dependent hyperadrenocorticism.

Dexamethasone Suppression, Canine/Feline Low­dose (Baseline, 4­hours, 8­hours)
Test Code: 127
Availability: Results available daily
Method: Chemiluminescence
Submission Requirements: 0.5mL serum (SST) per specimen
Storage: Refrigerate. Freeze if stored more than 3 days
Information: Sampling time: Baseline, 4­hour, and 8­hour post
Drug: Use either dexamethasone sodium phosphate or dexamethasone in polyethylene glycol.
Dosage: 0.01mg/kg IV
Interpretation:
Baseline cortisol: 1.0­5.0 ug/dL
4­hour and 8­hour post cortisol: < 1.4 ug/dL Canine: The 8­hour post sample is used to screen for hyperadrenocorticism: 8­hour post cortisol >1.4 ug/dL supports hyperadrenocorticism. The 4­hour post sample is used to help differentiate adrenal dependent versus pituitary dependent hyperadrenocorticism: 4hour cortisol < 1ug/dL supports pituitary dependent hyperadrenocorticism; If the 4­hour 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: Results available daily
Method: ELISA
Includes: Distemper and Parvovirus antibody
Submission Requirements: 1mL serum (SST)
Storage: Refrigerate for 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 HI antibody titer ≥1:80, indicating probable protective immunity. A negative result indicates a low antibody level, and a booster vaccination should be considered.

Ethylene Glycol (Qualitative)
Test Code: 345
Availability: Results available daily
Submission requirements: 2mL 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: Results available daily
Method: Guaiac test
Submission Requirements: 1 tablespoon fresh feces
Storage: Refrigerate
Suggested Protocol: Avoid red meat, (liver, beef, and lamb) and their by­products for 2­3 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 the 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
Method: IFA
Synonym: Feline Leukemia Virus
Submission Requirements: 2 air-dried UNSTAINED bone marrow or peripheral blood smears or 1mL whole blood (EDTA)
Storage: Refrigerate for up to 4 days, store slides at room temperature.
Inferences: Decreased platelets, decreased WBC count, poor slide quality, eosinophilia, slight hemolysis.
Interpretation: Positive or Negative. Vaccination does NOT interfere with this test

Fructosamine
Test Code: 354
Availability: Results available 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, and lipemia
Interpretation: This test reflects glycemic status over the previous 2­3 weeks.
The fructosamine concentration is directly related to the average blood glucose concentration in the preceding 2 to 3 weeks. The higher the average blood glucose concentration, the higher the serum fructosamine concentration. It may be helpful in differentiating elevated glucose values due to stress versus diabetes mellitus and in monitoring therapy for diabetes mellitus. Not affected by acute increases in blood glucose concentration that can occur with stress or excitement. Can be affected by concurrent hypoalbuminemia (mild decrease – dogs), hyperlipidemia (mild decrease – dogs), azotemia (mild decrease – dogs), hyperthyroidism (decreased – cats), hypothyroidism (decreased –dogs), prolonged storage at room temperature (decreased) and interfering substances such as hemolysis.
Patients Serum Fructosamine concentration Level of control

350 – 400 Excellent
400- 450 Good
450 – 500 Fair
>500 Poor
< 300 Prolonged hypoglycemia, diabetic remission (cat)

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. Culturettes 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 5­7 days
Method: AGID
Test includes: Blastomyces, Histoplasma, Coccidioidies and Aspergillus antibodies
Submission Requirements: 3mL serum (SST)
Storage: Refrigerate for up to 4 days
Interferences: Anticoagulants (EDTA or citrate) or preservatives
Interpretation: Positive or Negative; endpoint titer for Coccidioides­positive. 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 iron­binding 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. Patients affected by anemia of inflammatory disease will have a normal to low TIBC .

Lead Level
Test Code: 367
Availability: Results available in 3-5 days
Method: Atomic absorption
Submission Requirements: 1mL 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: Results available 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 ratio of citrate to whole blood, clotted specimen, non-citrate anticoagulant.

Phenobarbital
Test Code: 374
Availability: Results available 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: 15­40 ug/ml Possible toxic level: >40 ug/ml
The biological half­life of Phenobarbital in the dog ranges from 37–75 hours. The steady state is generally only achieved after 2­3 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: Results available within 24-36 hours
Submission Requirements: 1.5mL 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 pre­ovulatory 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: Results available daily
Submission Requirements: 1mL 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 2­3 days
Method: Latex Agglutination
Synonym: RF, rheumatoid arthritis
Submission Requirements: 1mL serum (SST)
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. Quantitative analysis gives the percent of each mineral present

T4, Total
Test Code: 387
Availability: Results available daily
Method: Chemiluminescence
Synonym: Thyroxine, tetraiodothyronine
Submission Requirements: 0.5mL 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 post­pill blood sample should be obtained 4­6 hours after thyroid supplement administration to monitor treatment efficacy.

Testosterone
Test Code: 818
Availability: Results available in 3-8 days
Method: RIA
Submission Requirements: 1mL serum (SST)
Storage: Refrigerate for up to 4 days
Interferences: EDTA
Interpretation:
Canine:
Male, intact: 1­7 ng/mL
Male, neutered: <0.1 ng/mL
Female: <0.1 ng/ml
Feline:
Male, intact: 1­6 ng/ml
Male, neutered: < 0.5 ng/ml
Female: <0.2 ng/ml
Equine:
Stallion, breeding season 1­4 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
Method: IFA
Submission Requirement: 1mL serum (SST)
Storage: Refrigerate for up to 4 days; Freeze for longer storage.
Inferences: 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 2­3 weeks.
* Test requires 24 hours incubation. Recommended test for canine and feline patients.

TSH, Endogenous, Canine
Test Code: 331
Availability: Results available daily
Submission Requirement: 1mL serum (SST)
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: Results available daily
Test Includes: Urine protein, Urine creatinine, Protein/Creatinine ratio
Normal Range: < 0.5 in dogs and < 0.4 in cats
Submission Requirements: 1mL urine in 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) or EDTA plasma (EDTA) 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: Freeze
Interferences: Hemoylsis, inclusion of RBC’s in plasma, or clotted specimen
* Plasma should not touch glass because it will cause clotting factor activation.
Interpretation: 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