Product: Anti-Spermatozoa Antibody (ASA) seminal ELISA
Anti-Spermatozoa Antibody (ASA) seminal ELISA
: 96 wells
: seminal plasma
: centrifugation, 1:5 dilution
: 100 µl
: 31 - 250 U/ml
: 1 h, 1 h (37°C), 30 min (RT)
: TMB at 450 nm
: 25 U/ml
The anti-spermatozoa antibody ELISA from Demeditec is a reliable and quantitative test for the determination antibodies directed against human spermatozoa. This test is intended for the use with seminal plasma.
Please note: the terms “anti-spermatozoa antibodies”, “anti-sperm antibodies” and “sperm antibodies” are equivalent. In these instructions the rather unwieldy but correct term “anti-spermatozoa antibodies” is used.
Antibodies directed against spermatozoa antigens may cause infertility in women or men. The application of the Anti-Spermatozoa Antibody ELISA from Demeditec is recommended for the diagnosis of immunologically caused disorders of fertility. Unwanted childlessness is a growing problem with which up to 20% of all couples in the reproductive age are confronted temporarily or long-term. In 20% of these cases the presence of anti-spermatozoa antibodies in the male or the female patient is detectable (Lahteenmaki A et al: Hum Reprod (1995) 10, 2824-28; Nagy ZP et al: Hum Reprod (1995) 10, 1775-80). The definition of infertility according to the WHO (WHO Laboratory Manual for the Examination of Human Semen and Semen Cervical-Mucus Interaction, 1999) is the absence of a conception within 12 months of unprotected intercourse. The main cause of an immunological fertility disorder is the formation of antibodies directed against spermatozoa antigens. Anti-spermatozoa antibodies exert heterogeneous effects on the ability of spermatozoa to fertilize. The inhibiting effect of anti-spermatozoa antibodies on the motility of spermatozoa by binding to their surface and by agglutinating processes is well-known (Zouari R et al: Fertil Steril (1993) 59, 606-12). The penetration of the spermatozoa into the cervical mucus is impaired by the presence of anti-spermatozoa antibodies in the seminal plasma and/or in the cervical mucus (Eggert-Kruse W et al: Hum Reprod (1993) 8, 1025-31). Anti-spermatozoa antibodies negatively influence the capacitation and the acrosome reaction of spermatozoa and thereby impede the interaction of the spermatozoa with the oocyte (Francavilla F et al: Front Biosci (1999): 1;4:9-25; Bohring C et al.: Hum Reprod (2001) 7:113-8). The interaction of the spermatozoon with the oocyte and the subsequent binding to and penetration of the zona pellucida may be inhibited by anti-spermatozoa antibodies. The following fusion of the oocyte and a spermatozoon may also be impaired by the presence of anti-spermatozoa antibodies (Mazumdar S et al.: Fertil Steril (1998) 70, 799-810; Kutteh WH: Hum Reprod, (1999) 14, 2426-9). According to Crosignani et al. (Crosignani et al.: PG et al.: Hum Reprod (1998) 13, 2025-32) the rate of pregnancies in couples with anti-spermatozoa antibodies on the part of the man or the woman are 38% lower compared to the control groups. Furthermore an influence on the implantation and on the early embryological development could be confirmed. An association of anti-spermatozoa antibodies and miscarriages is discussed. The frequency of anti-spermatozoa antibodies in infertile couples amounts to 20% (Lahteenmaki A et al.: Hum Reprod (1995) 10, 2824-28; Nagy ZP et al.: Hum Reprod (1995) 10, 1775-80). Anti-spermatozoa antibodies may occur dissolved in the ejaculate or bound to the surface of spermatozoa. Anti-spermatozoa antibodies may be found in men and in women (Clarke GN et al.: Am J Reprod Immunol Microbiol (1985) 7, 143-7). In women anti-spermatozoa antibodies may be found in cervical mucus, oviduct liquid and follicular liquid. Men having more than 50% of their spermatozoa coated with anti-spermatozoa antibodies show a conspicuously reduced rate of fertility (Abshagen K et al.: Fertil Steril (1998) 70, 355-6).
Fields of Application
The Anti-Spermatozoa Antibody ELISA from Demeditec can be applied in the clinical practice for the diagnosis immunologically caused infertility in men.
Principles of the Assay Method
The anti-spermatozoa antibody ELISA (Enzyme Linked ImmunoSorbent Assay) from Demeditec is a solid-phase sandwich enzyme-immunoassay for the quantitative determination of anti-spermatozoa antibodies in human seminal plasma. The ELISA-plate is coated with a mix of spermatozoa proteins which are recognized by anti-spermatozoa antibodies. The samples and standards are pipetted into the wells and then incubated. During this incubation anti-spermatozoa antibodies bind to the spermatozoa proteins and are thus immobilised on the plate. After washing the enzyme conjugate, consisting of anti-human globulin antibodies covalently coupled to horseradish peroxidase, is added. After removal of the unbound conjugate by washing the horseradish peroxidase oxidizes the then added substrate TMB (3,3’,5,5’-tetramethylbenzidine) yielding a colour reaction which is stopped with 0.5 N acidic solution. The extinction is measured at a wavelength of 450 nm with a microplate reader. The use of a reference measurement with a wavelength ³550 nm is recommended.