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The intra- and inter-assay variation of the indirect mixed antiglobulin reaction test: is a quality control suitable?
1Clinic of Dermatology, Department of Andrology, Clinical Training Center of the European Academy of Andrology, Philipp University, Deutschhausstrabetae 9, D-35033 Marburg, Germany.
Abstract:
The test most commonly used to detect sperm antibodies is the mixed antiglobulin reaction (MAR), standardized by the World Health Organization. The indirect MAR test detects soluble sperm antibodies in seminal plasma by using healthy donor spermatozoa as antigen. In this study we systematically investigated the influence of donor spermatozoa and the source of sperm antibodies upon the results of the indirect MAR test, and calculated the intra- and inter-assay variations. Using one individual seminal plasma and the same donor semen, results of the indirect MAR test are highly reproducible (low intra-assay variation). Two dimensions of inter-assay variation must be considered: (i) serial ejaculates of an individual donor may be used at different times; (ii) different donors may be applied to identical antibody sources. Donor spermatozoa strongly influenced the results of the indirect MAR test. Using multivariate statistical tests, highly significant main effects between the different donors (P < 0.001) and specific reciprocal effects between donor spermatozoa and seminal plasma samples (P < 0.001) were observed. The high inter-assay variation of the indirect MAR test will lead to incorrect results. There is urgent need of a reliable and reproducible test for sperm antibody detection to improve quality control of the methods.
Insights
The mixed antiglobulin reaction (MAR) test for sperm antibodies shows high variability due to donor sperm. This impacts test reliability, highlighting the need for improved sperm antibody detection methods.
Area of Science:
- Immunology
- Reproductive Medicine
- Clinical Diagnostics
Background:
- Sperm antibodies can affect male fertility.
- The mixed antiglobulin reaction (MAR) test is a common method for detecting sperm antibodies.
- The indirect MAR test uses donor spermatozoa as an antigen to detect antibodies in seminal plasma.
Purpose of the Study:
- To investigate the influence of donor spermatozoa and antibody source on indirect MAR test results.
- To determine the intra- and inter-assay variations of the indirect MAR test.
- To assess the reliability and reproducibility of the indirect MAR test for sperm antibody detection.
Main Methods:
- Systematic investigation of indirect MAR test variables.
- Use of multivariate statistical tests to analyze results.
- Calculation of intra- and inter-assay variations.
Main Results:
- Indirect MAR test results are reproducible with consistent seminal plasma and donor semen (low intra-assay variation).
- Significant variations were observed between different donors (P < 0.001).
- Specific reciprocal effects between donor spermatozoa and seminal plasma samples were highly significant (P < 0.001).
Conclusions:
- Donor spermatozoa significantly influence indirect MAR test outcomes.
- High inter-assay variation compromises the accuracy of sperm antibody detection.
- There is a critical need for a more reliable and reproducible test for sperm antibody detection to enhance quality control.
