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Progesterone Testing

We are available by phone Monday to Friday from 9:00 till 18:00.

For an appointment please call 06-25422015.
In case we are not available to pick up the phone please leave your name and number and a short message and we will return your call as soon as possible.

Outside these times and at the weekend we are available for artificial insemination and progesterone testing.

Our progesterone results are now available in 15 minutes! Immulite values.






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Frequently Asked Questions:

What is progesterone?

Progesterone is a hormone that is present in the female dog. It is produced in the ovaries. A characteristic of progesterone is that it increases in concentration as the bitch is ovulating. It’s concentration increases rapidly as the bitch approaches her ovulation peak.               

What is the purpose of a progesterone test?

The purpose of a progesterone test is to determine when the ovulation takes place. Without an ovulation the conception can not take place even though the semen of the stud dog is of excellent quality. “Timing” is the most essential factor.

“Timing” is of essence when:


● An old stud dog is used with a reduced libido ● A stud dog is used whose semen is of lesser quality ● A stud dog is used who has to mate 2 or more bitches in a short period of time. ● Frozen or chilled semen is used. ● The whelping date has to be determined in advance (dog breeds where caesarian are indicative) ● A bitch that shows little or no symptoms during the estrus. ● When the owner of the bitch has to drive a long distance to visit the stud dog. In such situations it is always pleasant to know that one is leaving at the correct time/date.


I have been informed that most bitches have to be bred on day 12 of their estrus cycle?

The average day of the ovulation peak is day 12 of the estrus cycle (3). One has to take into consideration that not every bitch follows the hormonal cycle according to the book! Research has shown that the ovulation peak can take place anywhere from day 7 until day 22 of the estrus cycle (3).   N B: It should be taken into consideration that not everyone takes into consideration as to when the estrus cycle of the bitch has taken place. Some bitches display little or no signs of estrus (vaginal discharge and interest in male dogs). It is therefore possible that a bitch in fact could be further with her estrus cycle then originally expected.

When should i start progesterone testing?

My advice to my clients is to start on day 7 of the estrus cycle. Depending on the results of the first blood results can a protocol be made with regards to further testing.

My bitch has already had 2 litters. During the last 2 matings she was mated on day 13 and 14 of the estrus cycle. Does it make sense to start progesterone testing again?

Progesterone results from the past are no guarantee with regards to the current estrus cycle.

The mother and grandmother of my bitch were bred on day 13 and day 14 of the estrus cycle and also became pregnant. Is there a genetic trait for this?

There is no scientific evidence that this is inherited.

If my bitch is bred with the correct progesterone results (advice from my veterinarian), does this guarantee that my bitch will also become pregnant?

Unfortunately there is no guarantee for this. A correct progesterone result (dependant on the type of test used) is only an indicator that the bitch has reached her ovulation peak. Other factors have to be taken into consideration such as the following:

● Harmful bacteria that can be located in the vagina and the uterus.
● The ovaries or the oviduct malfunction. ● Quality of the semen used ● Health condition of the bitch (illness)

How does the progesterone cycle work in the bitch?

The progesterone concentration begins to increase on average 2-3 days before that the ovulation peak is attained (2,3). At this point another hormone is released (Luteinizing hormone) which causes the ovulation to take place.  This unique hormonal encounter allows one to be able to determine when the bitch has to be bred.

Why is the Luteinizing hormone not measured instead of the progesterone concentration?

It is more difficult to measure the Luteinizing hormone in comparison to that of progesterone. The Luteinizing hormone release is also of a short period (1-2 days)(2,3).


The advantage of measuring the progesterone concentration is that it’s increase can be measured gradually throughout the estrus cycle. The ovulation peak occurs 2-3 days after the Lutenizing hormone peak, and the conception can take place 4-6 days after the Lutenizing hormone peak has taken place (3).

Which types of progesterone tests are available on the market?

There are many types of progesterone tests available. Most of the available tests are based on that of radioimmunossay (RIA) or that of an enzyme-linked immusorbent assay (ELISA).


RIA test is more accurate than that of the ELISA (6) but also more expensive. The test is based on a quantitative result.

The ELISA test can be both quantitative as well as semi-quantitative. With some ELIZA tests the colour of the progesterone concentration of the bitch is compared to that of the reference values. Depending on the colour displayed can a progesterone value be determined.

Are there other methods available to help determine the correct time to breed a bitch?

Another method is to take a vaginal smear and to examine this under the microscope (1). During this examination the changes in the cell structure (epithelial cells) are examined. Depending on the changes can an estimate be made as to when to breed the bitch.

My bitch is now in estrus (day 1). What should I do?

There are different progesterone tests available on the market. Each manufacturer has it’s own protocol. I shall give a short summary as to how I determine progesterone based on the test method that I use in my clinic.




On day 1 of the estrus cycle I advise the owner of the bitch to have bacterial examination done of the reproductive tract to determine if there are any bacteria that can have a negative influence on the fertility. The owner of the bitch comes to the clinic with his bitch on day 7 of her estrus cycle. A blood sample is taken (1-2 ml). This blood sample is placed in a centrifuge machine from which the plasma is separated. Afterwards the plasma is removed and used for the progesterone test.


Four different standard concentrations of progesterone are used as reference points (1.0, 2.5, 5.0 and 10.0 ng/ml progesterone). From the plasma attained, 2 samples are used. Afterwards chemical components are added to all 6 samples (according to a protocol).

After approximately 1 hour the samples are placed into a special machine that measures the light intensity/refractometry. The tabulated results are printed out afterwards. With the use of a graph can the progesterone concentration be measured. This test is based on a quantitative assay.


Depending on the result is it possible to give advice as to when another test should be done or to advice the owner as to when to have their bitch mated.


Guidelines that I use with this test method is as follows:

  • < 3 ng =  retest in 3 days
  • 5-7 ng = retest in 2 days
  • 7-10 ng = breed within 24 hours
  • >10 ng = breed directly


Fogle, B. Caring For Your Dog. Dorling Kindersley Ltd. Great Britain. 2002 p. 52 – 56.


Hase et al. Plasma LH and progesterone levels before and after ovulation and observation of ovarian follicles by ultrasonographic diagnosis system in dogs. J Vet Med Sci. 2000 Mar;62(3):243-8.


Johnston, S. et al. Canine and Feline Theriogenology. Saunders, An Imprint of Elsevier U.S.A. 2001. P. 49-52, 50, 52, 56-57.


Soderberg, S. Canine breeding management. Vet Clin North Am Small Anim Pract. 1986 May;16(3):419-33.


5.= Tilley, L. et al. The 5 minute Veterinary Consult. Lippincott Williams & Wilkins U.S.A. 2004 p. 706 – 707.


Van Klaveren, NJ et al.The optimal mating time in the bitch based on the progesterone concentration in peripheral blood. A comparison of reliability between three ELISA test kits and a 125-iodine radioimmunoassay. Tijdschrift Diergeneeskd. 2001 Nov 1;126(21):680-5. Dutch.