Spermatozoa Ab ELISA Kit

Sample

Serum

Species Reactivity

Human

Detection Method

sELISA

Intended Use

The anti-spermatozoa antibody ELISA is a reliable and quantitative test for the determination antibodies directed against human spermatozoa. This test is intended for the use with serum. This assay kit is for research use only and not for use in diagnostic or therapeutic procedures.

Contents of Kit

1. Microtiter strips coated with sperm antigen 96 wells
2. Sperm Antibody ELISA standard set - per vial 0.5 ml
– Standard 1 ( 31 U/ml – colourless screw cap)
– Standard 2 ( 62 U/ml – white screw cap)
– Standard 3 (125 U/ml – yellow screw cap)
– Standard 4 (250 U/ml – blue screw cap) 0.5 ml
3. Control (green screw cap) 0.5 ml
4. Dilution buffer (also used as blank / zero standard / 0 U/ml ) 50 ml
5. Washing solution (10× concentrated) 50 ml
6. Enzyme conjugate (ready for use) 8 ml
7. Substrate solution (solution of TMB, ready for use) 13 ml
8. Stop solution (0.5 N/l acidic solution) 13 ml
9. Holder for single strips 1 ×

Storage

1. Store the reagents at 2°C-8°C.
2. The reagents remain stable until the expiration date of the kit.
3. The diluted washing solution is stable for 4 weeks at refrigerator temperatures (4°C-8°C).
4. Put caps back on the vials immediately after use.
5. Store the microtiter strips in a dry bag with desiccants. The remaining strips must be stored in the tightly resealed bag together with the desiccants. Under these storage conditions, they are stable at least for 4 weeks after opening of the sealed bag.

Performance Characteristics

Expected Values
Normal values 0 – 60 U/ml
Elevated values above 60 U/ml
In the case of a value in the range near the cut-off (55 to 65 U/ml) we recommend a follow-up determination using a new sample taken within the next two weeks.

Precision

1. Intraassay variation coefficient: 6.88% (5.90-7.81 %)
For the determination of the intraassay variation coefficient 6 kits from 6 different batches (produced on different days) were used. One patient sample (optical density about 1.0) was applied 96 times per testing procedure.
2. Interassay variation coefficient: 6.45% (4.84-7.52 %)
For the determination of the interassay variation coefficient one strip each of 12 kits stemming from 6 different batches (produced on different days) were used. One patient sample (optical density about 1.0) was applied 72 times per testing procedure.

General Description

Antibodies directed against spermatozoa antigens may cause infertility in women or men. The application of the Anti-Spermatozoa Antibody ELISA 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).