Test Protocol Index
Test Protocol 11

The ACTH stimulation test is indicated as a screening test for hyperadrenocorticism, and as a confirmatory test for hypoadrenocorticism and iatrogenic hyperadrenocorticism in dogs. It may also be used to monitor Mitotane therapy in hyperadrenocorticoid dogs (Dunn et al, 1995).


  1. Take a basal sample of 2ml clotted or heparinised blood after an overnight fast. (Taking the basal sample at 9 to 10 a.m. will minimise the influence of diurnal fluctuations in cortisol concentration).
  2. Inject 0.25mg synthetic ACTH i/v (Synacthen).
  3. Take a second 2ml of clotted or heparinised blood 1 hour post Synacthen injection.
  4. Separate the serum or plasma before despatch to the laboratory and clearly label the tubes "pre" and "post" ACTH.

An increase in cortisol levels post ACTH is expected in all dogs. In normal dogs levels will rise to less than 660 nmol/l. A post ACTH cortisol level of >660 nmol/L is POSITIVE for hyperadrenocorticism Dogs with pituitary dependant hyperadrenocorticism (PDHAC) show an exaggerated response usually greater than 660 nmol/l. In dogs with an adrenal tumour the basal cortisol is usually high with a variable response to ACTH stimulation. In cases of Addisons Disease (primary hypoadrenocorticism) or iatrogenic hyperadrenocorticism, basal cortisol values are normally low with little or no response to ACTH stimulation.


The ACTH stimulation test will identify approximately 85% of PDHAC dogs and approximately 50% of dogs with an adrenal tumour. It will not reliably distinguish between the two conditions. The dexamethasone (high dose) suppression test is recommended for distinguishing between PDHAC and an adrenal tumour following a positive ACTH stimulation test result


Please enquire.
Test Code Test Name and Description Sample
ACTH ACTH Stimulation Test (Canine) 2 x serum or heparin plasma


Evans H and Walker M. SCL Veterinary Services Manual. 6th Edition. p.15.

Dunn, K.J., Herrtage, M.E. and Dunn, J.K. (1995) Use of ACTH stimulation tests to monitor the treatment of canine hyperadrenocorticism. Veterinary Record 137(7) p 161 - 165.

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