Human Sperm-Antibody ELISA Kit

Sample

Serum

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The Human Sperm-Antibody ELISA is an enzyme immunoassay for the measurement of antibodies directed against human spermatozoa in serum. It is intended for research use only.
The device is intended to be used as an aid for the diagnosis of immunologically caused disorders of fertility;
The device is not intended to be used for the detection of poly- and monoclonal gammopathies.

Contents of Kit

1. Microtiters strips coated with sperm antigen, 96 wells
2. CAL N Calibrators, N=1 to 4
4 vials, 0.5 ml, ready to use
Calibrator 1 (31 U/ml)
Calibrator 2 (62 U/ml)
Calibrator 3 (125 U/ml)
Calibrator 4 (250 U/ml)
3. Ab HRP Enzyme Conjugate, 1 vial, 8 ml
Anti-human IgG antibody conjugated with horseradish peroxidase, ready to use
4. DIL BUF
1 vial, 50 ml/vial, ready to use
Also used as blank / zero calibrator 0 U/ml
5. CHROM TMB
1 vial, 14 ml
Ready to use
Solution of TMB
6. STOP SOLN
1 vial, 14 ml, ready to use
Contains 0.5M H₂SO₄
7. WASH SOLN CONC
1 vial, 30 ml (40× concentrated)
8. CONTROL N
N=1, 1 vial, 0.5 ml, ready to use

Storage

When stored at 2 °C to 8 °C unopened reagents will retain reactivity until expiration date. Do not use reagents beyond this date. Opened reagents must be stored at 2 °C to 8 °C. Microtiter wells must be stored at 2 °C to 8 °C. Once the foil bag has been opened, care should be taken to close it tightly again. Opened kits retain activity for 8 weeks if stored as described above.

Precision

Within-run Precision
The within-run precision was determined with 4 patient samples covering the complete measuring range within 20 days in 2 independent runs per day. CV was calculated as mean CV of 40 runs.

Between-run Precision
The between-run precision was determined with 4 patient samples covering the complete measuring range within 20 days in 2 independent runs per day and with 2 replicates per run (20 x 2 x 2). CV was calculated from 80 determinations.

Between-lot Precision
The between-lot variation was determined by 6 measurements of different samples with 3 different kit lots.

Detection Range

Measuring range: 3.367 U/mL - 250 U/mL
Linear range: 13.333 mL - 250 U/mL

Sensitivity

Limit of Blank (LoB): 0.490 U/mL
Limit of Detection (LoD): 3.367 U/mL
Limit of Quantification (LoQ): 9.632 U/mL

General Description

Antibodies directed against spermatozoa antigens may cause infertility in women or men. The application of the Sperm 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 5-20 % of these cases the presence of anti-spermatozoa antibodies in the male or the female patient is detectable.
The definition of infertility according to the WHO (WHO Laboratory Manual for the Examination of Human Semen and Semen CervicalMucus 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 (ASA) exert heterogeneous effects on the ability of spermatozoa to fertilize. The inhibiting effect of ASA on the motility of spermatozoa by binding to their surface and by agglutinating processes is well-known.
The penetration of the spermatozoa into the cervical mucus is impaired by the presence of ASA in the seminal plasma and/or in the cervical mucus. ASA negatively influence the capacitation and the acrosome reaction of spermatozoa and thereby impede the interaction of the spermatozoa with the oocyte.
The interaction of the spermatozoon with the oocyte and the subsequent binding to and penetration of the zona pellucida may be inhibited by ASA. The following fusion of the oocyte and a spermatozoon may also be impaired by the presence of ASA.
The rate of pregnancies in couples with ASA on the part of the man or the woman was shown to be 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 ASA and miscarriages is discussed. The frequency of ASA in infertile couples amounts to 20%.
ASA may occur dissolved in the ejaculate or bound to the surface of spermatozoa. ASA may be found in men and in women. In women, ASA 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. ASA have been shown to be associated with chronic prostatitis which has a negative effect on male reproductive function.

Standard Curve

The following data is for demonstration only and cannot be used in place of data generations at the time of assay.