Frequently Asked Questions: SpermMar IgG

FAQ1: How long can we wait before the test should be performed?
Perform the test preferably within 1h after collection, especially in samples with low motility. If motility is high, it might be possible to perform the test later (max 3-4h later).

FAQ2: Can we perform the SpermMar IgG test on washed spermcells ?
We advice against washing the spermcells before performing the direct SpermMar IgG test for a number of reasons.
First of all, some washing media contain human serum albumin (HSA). HSA tends to form a “shield” around the spermcells which hide the antibodies so that they can not be detected by the test method.
Secondly, washing the sperm sample will remove most of the antibodies that could be present in the seminal plasma and might thus render false negative results.

FAQ3: We do the SpermMar IgG test, should we also do the SpermMar IgA test ?
Most laboratories only do the SpermMar IgG on a routine basis, in case of a positive IgG they also do an IgA.
It does happen however that a patient with a negative IgG is positive for IgA. So, from a clinical point of view you should do both SpermMar IgG and SpermMar IgA, from an economical point of view however you could probably opt for the IgG as a routine test and IgA for positive IgG’s.

FAQ4: Why does the SpermMar IgG kit contain 2 bottles and the SpermMar IgA only 1 ?
The SpermMar IgG kit contains one bottle of latex particles coated with IgG and one bottle of anti-IgG antiserum. The antiserum actually links the antibodies on the surface of the spermatozoa to the antibodies on the surface of the latex particles.
The SpermMar IgA on the other hand contains only one bottle of latex particles coated with monoclonal anti-IgA directly linking the latex particles to the antibodies on the surface of the spermatozoa. The SpermMar IgA therefore only needs 1 bottle of reagents where the SpermMar IgG needs 2.

FAQ5: Sometimes when I do the SpermMar IgG test, the spermcells stop moving after 15 minutes, is this normal ?
Yes it is, after mixing the reagents with the spermatozoa wait 2-3 minutes before reading the results, you should not wait longer than 5 minutes.

FAQ6: In the SpermMar IgG test the latex particels agglutinate, is this normal ?
Yes it is, because of the principle explained in faq3, the antiserum not only links the latex particels to the spermatozoa but also to the other latex particels which also contain IgG. The particels therefore tend to agglutinate.
Another reason why agglutination may occur is because the bottles containing the latex particles was not mixed well before use. Also make sure to thoroughly mix the antiserum with the latex particels and the sperm on the slide.

FAQ7: What is the origin of the antibodies and other biological material?
The SpermMar Test IgG contains bovine sourced albumin. The beads are coated with human IgG antibodies, and its antiserum contains rabbit anti-human IgG antibodies.

FAQ8: Do you supply positive or negative controls for the SpermMar IgG test ?
Yes we do, the controls are already diluted by us and can be used directly out of the bottle. The controls can only be used in the indirect SpermMar test.
Note: The positive and negative controls are made up of human sourced materials and should be treated as potentially infectious.

FAQ9: When is it recommended to perform a quality control? How do we perform quality control on the SpermMar Test IgG?
This decision is lab-dependent. Most labs do a quality control when starting a new lot. This lot can be tested with the SpermMar Test IgG Positive and Negative Control. The results should be ≥80% and <40% respectively.

FAQ10: What is the difference between the single kit and the complete kit?
The single kit only contains the reagents, while the complete kit also contains the lab disposables (micro slides, cover glasses, microcapillary pipettes with rubber bulb). The complete kits do not contain the controls.

FAQ11: What are the minimal spermatozoa parameters needed to perform a direct test? When is an indirect test recommended?
A concentration of >10 million/ml and a progressive motility of >30 % is advisable. Otherwise, the indirect test is a more suitable option. The sperm samples can be concentrated or diluted, but it is advisable not to use a medium containing HSA or another protein supplement as this can interfere with the test results.

FAQ12: Can we perform the Indirect SpermMar IgG test on frozen/thawed semen plasma samples?
Yes it can. It is best centrifuge the UNTREATED semen sample before freezing the semen plasma at -18C or below.

FAQ13: Can the direct test be done on frozen/thawed semen samples?
No, it is recommended on native samples because HSA in cryoprotectant can mask anti-sperm antibodies on spermatozoa.

FAQ14: What is the shelf life after opening?
If the product is used aseptically, firmly closed and kept refrigerated between tests, the shelf life after opening remains the same as when unopened (18 months).