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Frequently Asked Questions


Does it matter which collection tubes are used for test sample? Do tubes with serum separaters affect test results?
 

The color test result is not uniformly round (it may be a crescent or half moon shape).  

I have misplaced the pipettes in the kit. How much sample do I need?

Target Test result is consistently blue though white result is expected.  

Can Target Canine Test be used to detect pregnancy in the dog?

Target Test result is consistently white though the bitch is showing all the other signs of estrus.

How is the Target canine test kit used for timing a C-Section?

What is the minimum amount of sample that can be used for a test?  

Can a hemolyzed serum sample be used for the test?  

Can I check to see if the test reagents are working correctly?

Can a pregnant mare show signs of estrus?  

Can I freeze milk samples to test at a later date?

 

 

 

Q. Does it matter which collection tubes are used for test sample? Do tubes with serum separaters affect test results?

A. Serum is the sample of choice for the Target canine progesterone kit, and plain Red Top blood collection tubes are preferred for serum collection. This removes any danger of interference of test results from chemicals used in coating other tubes. Also, as blood collected in Red Top tubes clots at the bottom of the tube with the serum rising to the top, the serum can be easily siphoned off onto the test cup with the fine-tipped pipettes provided in the kit. You do not need a centrifuge to run this test.
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Q. The color test result is not uniformly round (it may be a crescent or half moon shape).

A. The filter test cup may be partially clogged by the sample. Repeat test using better quality sample.

Sample Preparation: The quality of a sample affects the accuracy of any test. Since the Target test cup is a filter, it is especially important that the sample does not contain a large amount of precipitate or red blood cells which can cause the filter to be clogged and lead to inaccurate results.

Poor Quality Samples: If a sample looks unusually hemolyzed (red), is lipemic, or looks especially viscous, repeat the wash step 2 several times to make sure that the sample debris is washed away before adding the enzyme in step 3. Use caution in interpreting the result.
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Q. I have misplaced the pipettes in the kit. How much sample do I need?

A. Enough sample to cover the top surface of the test cup (to ensure binding with antibody at all sites). This is about 3-4 drops (150-200 micro liters) of sample. The size of sample is not important, since the sample is always in excess over the amount of antibody present on the test cup. Too much sample does not adversely affect test result, but not enough sample to cover the top surface of the test cup does).
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Q. Target Test result is consistently blue though white result is expected.

A. Sometimes high progesterone agglutinates or clumps together in a lump at one site, making it difficult for the test to find progesterone to bind to at other sites. Also, the test is designed for ovulation timing, when progesterone is low, so in rare cases extremely high levels of progesterone could result a false reading. Because Target is a rapid test, the technology does not allow for specific numeric value. Send sample to a lab for RIA to get a numeric value for the level of progesterone.
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Q. Can Target Canine Test be used to detect pregnancy in the dog?

A. No, the Target canine test is for ovulation timing, not pregnancy detection. There is no difference of diagnostic value between progesterone from the pregnant and the nonpregnant bitch because of the unique characteristic of the bitch where the corpus luteum, (from which progesterone is made), is functional for the same length of time in the nonpregnant and the pregnant bitch. Progesterone concentrations peak approximately 20 to 25 days following ovulation at 30-60 ng/ml, followed by a transient plateau and a prolonged decline until progesterone is less than 1 ng/ml. The hormone relaxin is a marker for pregnancy in the dog because serum relaxin is low in nonpregnant bitches and rises significantly in pregnant bitches after Day 21.
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Q. Target Test result is consistently white though the bitch is showing all the other signs of estrus.

A. Run a control on the test by following protocol starting at Step 2 (i.e. do not add any sample) The result should be a bright blue. If it is, then the problem might be with the quality of the sample or the health of the dog. (Luteal cysts will secrete progesterone, and may cause the test result of a dog in heat to be white). If the Control test result is white, call Target at 1-800-999-1961.
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Q. How is the Target canine test kit used for timing a C-Section?

A. Approximately 58-68 days post breeding, when progesterone drops to bright blue (C1, or close to it) after the previous test was light blue or close to white, C-Section should be scheduled to take place within 24 hours.

During pregnancy progesterone is present at high levels to maintain fetus viability. Progesterone is highest at 30-60 ng/ml around day 25. It declines very slowly until about 3 days pre-partum to around 5ng/ml. Then about 24 hours before birth, there is a dramatic drop of progesterone levels to less than 2 ng/ml. The color of the Target test changes from about C4 to about C1.

Premature puppies lack surfactant which prevents their lungs from “sticking” together, allows their lungs to inflate and deflate with every breath taken. Studies show that when progesterone falls to less than 2 ng/ml, surfactant has been released, and the pregnancy is at its natural end. Puppies born will therefore be viable. The stimulation of surfactant production is also an effect of prepartum rise in fetal glucocorticoids which takes place at this time as progesterone declines.

Other observations about the bitch's condition should be noted and used in conjunction with progesterone test. C-sections undertaken too early may result in puppies that are not viable.

Other signs to look for:

•  drop in rectal temperature to below 100oF (normal Temp 101-102.5 0F
•  restlessness with panting; uneasiness
•  seeking of quiet place to build nest
•  mucus plug being discharged

Has it been between 58-68 days since breeding? Average gestation is 63 days, though smaller breeds generally tend to whelp sooner at around 60 days. Bulldogs and Boston Terriers almost always need to be delivered by C-Section, and sooner rather than later at around 58-59 days. Other breeds that often require C-section when whelping are Pugs, Chihuahuas and Pekingese.
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Q. What is the minimum amount of sample that can be used for a test?

A. The 8 drops suggested in the instructions insures that the top of the test cup has been fully covered. However, by visual inspection, you can see that 3 or 4 drops will cover the surface and could be the minimum amount of sample used.
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Q. Can a hemolyzed serum sample be used for the test?

A. Yes, a hemolyzed sample can be used. In Step 2 of the instructions when sample wash is added to the cup, any remaining red blood cells will be lysed and the sample will be washed. A centrifuge is helpful for sample preparation; but not required.
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Q. Can I check to see if the test reagents are working correctly?

A. Yes, run a test starting with Step 2 of the instructions. When no sample has been added, the test result will be bright blue (C1 result) since no progesterone has been added.
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Q. Can a pregnant mare show signs of estrus?

A. Yes, about 30% of all mares show heat when they are actually in foal. In the mare, an unusual situation prevails leading to follicle growth and even ovulation early in pregnancy. (Follicle growth is generally depressed though during pregnancy because of the accompanying high levels of progesterone).
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Q. Can I freeze milk samples to test at a later date?

A. No, it is best to test milk as soon as possible. Once milk is frozen, it never regains a homogenous nature when thawed. The fat and water portions separate out like oil and vinegar. Since progesterone binds mainly to fat and not water, test results from thawed milk sample will not be representative of progesterone levels in the sample as a whole. If not tested within 30 minutes of collection, milk sample should be refrigerated. A preservative, potassium dichromate, should be added to sample if testing will not be done within 6 hours. Also bromothymol can be used.
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