Sperm-cervical Mucus Penetration Test Research Articles

Temporal changes in cervical mucus after insertion of the levonorgestrel-releasing intrauterine system

BackgroundThe major contraceptive action of the levonorgestrel-releasing intrauterine system (LNG-IUS) is cervical mucus (CM) thickening, which prevents sperm penetration. No study to date has examined the temporal relationship between the insertion of the LNG-IUS and changes in CM quality and sperm penetration. Study DesignParticipants were enrolled in a clinically descriptive study to compare the quality of CM and three parameters of sperm penetration prior to insertion of the LNG-IUS and on Days 1, 3 and 5 after insertion. Measurements of estradiol, progesterone and levonorgestrel (LNG) in serum and LNG in CM were also carried out at these times. CM was analyzed using the World Health Organization CM grading criteria. Sperm penetration was determined using an in vitro sperm–CM penetration test. ResultsAll 10 participants underwent LNG-IUS insertion during midcycle when CM quality was good and sperm penetration was excellent. On Day 1 after LNG-IUS insertion, the majority of participants demonstrated poor CM quality and poor sperm penetration. On Day 3, all participants had poor CM quality, and all but one subject had poor sperm penetration. By Day 5, all participants had poor CM quality and poor sperm penetration. LNG levels in CM peaked on the day after LNG-IUS insertion. ConclusionSignificant changes in quality of CM and sperm penetration were observed shortly after LNG-IUS insertion; however, CM can remain penetrable for up to 5 days when the LNG-IUS is inserted midcycle.

Effects of the levonorgestrel-releasing intrauterine system on cervical mucus quality and sperm penetrability

Background In levonorgestrel intrauterine system (LNG-IUS) users, the prevention of sperm penetration through cervical mucus has not been demonstrated. Study Design Subjects were enrolled in an investigator-blinded study to compare quality and sperm penetrability of mid-cycle cervical mucus between LNG-IUS users and hormone-free controls. Cervical mucus was microscopically examined using World Health Organization (WHO) cervical mucus analysis (CMA). CMA score ≥10 of 15 points indicated cervical mucus favoring sperm penetration. Mucus was incubated with sperm using the WHO simplified slide test (SST) and Kremer sperm cervical mucus penetration test (SCMPT). Results Data from 14 LNG-IUS users and 16 controls showed 14% of LNG-IUS users had CMA score ≥10% vs. 69% of controls (p=.004). SST showed no sperm penetration for LNG-IUS users, significantly less than controls (0% vs. 64.3%, p<.001). SCMPT demonstrated no sperm mucus penetration for LNG-IUS users at 2 and 6 h (0% vs. 85% in controls with 2-h score ≥6, p<.001; 6 h 0% vs. 79% in controls, p<.001). Conclusions Mid-cycle cervical mucus of LNG-IUS users is poor quality and prevents endocervical sperm transport in vitro.

Effects of the levonorgestrel intrauterine system (LNG-IUS) on cervical mucus quality and sperm penetration

OBJECTIVE: No study has shown that changes in cervical mucus prevent sperm penetration in LNG-IUS users. The purpose of this study was to compare the depth of sperm penetration in the mid-cycle cervical mucus of LNG-IUS users to non-users with in-vitro sperm penetration tests using standardized sperm from a single donor; and by performing WHO cervical mucus grading.DESIGN: 20 LNG-IUS users and 20 controls are being enrolled in an investigator-blinded, age-matched, pharmaco-physiologic study.MATERIALS AND METHODS: Cervical mucus assessment at mid-cycle was estimated by home urinary luteinizing hormone tests. Cervical mucus was microscopically examined and graded using WHO criteria. Mucus was incubated with sperm from one standardized donor using both the Kremer Sperm Cervical Mucus Penetration Test (SCMPT) and the WHO Modified Slide Test. Depth of linear penetration of the vanguard sperm, density of penetration and forward motility were assessed microscopically and categorically graded. All parameters were dichotomized and compared using Fisher Chi-Square tests.RESULTS: To date, 46 subjects have been enrolled. Twelve LNG-IUS users and fifteen controls have completed the study. Of users, 20% had cervical mucus analysis grading over 10 (“good”) vs. 71% of controls (p=.02). WHO Modified slide test showed significantly less mucus with penetration by sperm for LNG-IUS users than controls (11% vs. 58%, p=.04). SCMPT demonstrated significantly lower “good/excellent” penetration of cervical mucus by sperm for users vs. controls at 2 and 6 hrs (0% vs. 85% at 2 h, p =.0005; 0% vs 77% at 6 h, p =.002). No cervical mucus from LNG-IUS users had good sperm penetration of mucus.CONCLUSIONS: The quality and sperm penetrability of mid-cycle cervical mucus are significantly less among LNG-IUS users than controls. The high contraceptive efficacy of the LNG-IUS occurs because the local release of levonorgestrel prevents sperm transport through the cervical mucus to the endometrial cavity, and fertilization does not occur. OBJECTIVE: No study has shown that changes in cervical mucus prevent sperm penetration in LNG-IUS users. The purpose of this study was to compare the depth of sperm penetration in the mid-cycle cervical mucus of LNG-IUS users to non-users with in-vitro sperm penetration tests using standardized sperm from a single donor; and by performing WHO cervical mucus grading. DESIGN: 20 LNG-IUS users and 20 controls are being enrolled in an investigator-blinded, age-matched, pharmaco-physiologic study. MATERIALS AND METHODS: Cervical mucus assessment at mid-cycle was estimated by home urinary luteinizing hormone tests. Cervical mucus was microscopically examined and graded using WHO criteria. Mucus was incubated with sperm from one standardized donor using both the Kremer Sperm Cervical Mucus Penetration Test (SCMPT) and the WHO Modified Slide Test. Depth of linear penetration of the vanguard sperm, density of penetration and forward motility were assessed microscopically and categorically graded. All parameters were dichotomized and compared using Fisher Chi-Square tests. RESULTS: To date, 46 subjects have been enrolled. Twelve LNG-IUS users and fifteen controls have completed the study. Of users, 20% had cervical mucus analysis grading over 10 (“good”) vs. 71% of controls (p=.02). WHO Modified slide test showed significantly less mucus with penetration by sperm for LNG-IUS users than controls (11% vs. 58%, p=.04). SCMPT demonstrated significantly lower “good/excellent” penetration of cervical mucus by sperm for users vs. controls at 2 and 6 hrs (0% vs. 85% at 2 h, p =.0005; 0% vs 77% at 6 h, p =.002). No cervical mucus from LNG-IUS users had good sperm penetration of mucus. CONCLUSIONS: The quality and sperm penetrability of mid-cycle cervical mucus are significantly less among LNG-IUS users than controls. The high contraceptive efficacy of the LNG-IUS occurs because the local release of levonorgestrel prevents sperm transport through the cervical mucus to the endometrial cavity, and fertilization does not occur.

The effect of heavy metals on the in vitro interaction between human sperm and cervical mucus

The sperm-cervical mucus-penetration test (SCMPT) was used in vitro to examine the influence of heavy metals (mercury, cadmium and copper) on sperm function. 30 each of freshly obtained different sperm samples with varying starting sperm motility and oestrogen-stimulated cervical mucosa from women of known fertility were used. Mercury chloride (HgCl2; 0.005-0.5 mmol/l), cadmium chloride (CdCl2; 0.05-10 mmol/l) and copper sulphate (CuSO4; 0.0001-10 mmol/l) were added to the in-vitro system. After incubation, at 37 degrees C for 6 hours, linear progression, penetration depth and motility index of the sperm (cumulative SCMPT score) were analysed. An untreated control was run in parallel with each test. The added substances, especially CdCl2 and HgCl2, significantly reduced the quantitative and qualitative penetrability of sperm (P = 0.001), in step with the concentration. The concentrations which were used, especially those of mercury, were in the range of in-vivo. measured levels. These results make clear that heavy metals can influence human sperm function.

Prevalence of Chlamydia trachomatis in subfertile couples

Prevalence of Chlamydia trachomatis in subfertile couples

Análisis del factor cervical de esterilidad

To investigate the role of both cervical mucus and semen parameters in diagnostic tests of the cervical factor: Insler test, postcoital test (PCT) and in vitro sperm-cervical mucus penetration test (PT), with special interest in identified pathogenic organisms. We investigated infertility in 376 infertile patients, concentrating on the cervical factor in 281 of them (74.73%), and using three test methods. Insler (n = 201), postcoital (n = 254), and PT (n = 79), and endocervical and vaginal secretion cultures in all of them. Also the direct microimmuno-fluorescence test (Micro-Trak) ® and 222 routine sperm tests on their partners. Insler test was normal in 72.6% of the cases, without significant (NS) correlation with infection. The postcoital test was pathological in 72.55% of cases. We correlated these with the Insler test results ( P < 0.005; χ 2 1 = 105.670), TPV results (NS), semen parameters ( P < 0.005; χ 2 1 = 4.441) and infection (NS), particularly with Chlamydia trachomatis (χ 2 1 = 0.529; P < 0.005). The PT showed in 45.8% of cases the sperm did not penetrate, in 24.1% it did, in 3.6% mobility was lowered, and in 26.5% it was immobile, noting that PT was in correlation with sperm analysis ( P < 0.005; χ 2 1 = 3.920). The results of the postcoital test depend on the properties of the mucus and semen, these being altered by the presence of C. trachomatis infection, so that PT is related to the sperm parameters. Se ha evaluado la influencia tanto del moco cervical como de las características del semen en las diversas pruebas diagnósticas del factor cervical: test de Insler,test poscoital y test de penetración in vitro ,con especial atención al papel que desempeñan los microorganismos patógenos identificados. Se efectuó el protocolo de esterilidad a un total de 376 mujeres,centrándonos en el análisis del factor cervical,llevado a cabo en 281 de ellas (74,73%), con tres pruebas:de Insler (n = 201), poscoital (n = 254) y de penetración in vitro (n = 79),además de cultivos de la secreción endocervical y vaginal a todas ellas, y test de microinmunofluorescencia directa para la identificación de Chlamy dia trachomatis (Micro-Trak ® ),así como 222 espermiogramas a sus parejas. El test de Insler fue normal en el 72,6% de los casos,no hallando relación con la infección (NS). El test poscoital se estableció como patoló gicoen el 72,5% de los casos. Se relacionaron sus resultados con el test de Insler (p < 0,005; χ 2 1 = 5,670), el test de penetración in vitro (NS),con los del espermiograma (p < 0,005; χ 2 1 = 4,441) y con la infección demanera global (NS) y por C. trachomatisen particular(χ 2 1 = 0,529; p < 0,005). En el test de penetración in vitro se observó que en un 45,8% de los casos los espermatozoides no penetraban,en un 24,1% sí lo hacían,en tanto que en un 3,6% se disminuía su movilidad y en un 26,5% se inmovilizaban,observando que el test de penetración in vitro se relacionaba con el espermiograma (p < 0,005:χ 2 1 = 3,920). Los resultados del test poscoital dependen de las características del moco y del semen, y se encuentran alterados ante la presencia de infección por C. trachomatis ,en tanto que el test de penetración in vitro se relaciona con los valores del espermiograma.

Sperm morphology assessment using strict criteria and male fertility under in-vivo conditions of conception

The clinical significance of sperm morphology assessment according to very strict criteria was determined using semen samples of randomly chosen males from couples not submitted to assist procreation techniques, with a median duration of infertility of 4 years (range 1-17; n = 89). The relationships of sperm morphological properties to the results of standard sperm analysis, including the differentiation of round cells in semen by monoclonal antibodies and semen cultures, the testing of sperm functional capacity in vitro with the standardized sperm-cervical mucus penetration test (SCMPT) and the subsequent pregnancy rate under in-vivo conditions of conception, were evaluated in a prospective study. The quick staining method (DiffQuick(R) stain) for sperm morphology proved to be practical and suitable for routine use. The percentage of normal forms according to strict criteria ranged from 1 to 36%, with a median of 12%. Morphological findings were not markedly related to the medical history, but significant relationships between standard parameters of sperm analysis, in particular the sperm count, the progressive motility and standard sperm morphology, were found. The percentage of normal forms was not significantly associated with the microbial colonization of semen samples but was negatively related to high leukocyte rates. Semen samples with a higher percentage of normal spermatozoa (shown, for example, for >4,> 7 or >=14% normal) were significantly more frequent in cases of an adequate SCMPT. The subsequent pregnancy rate within an observation period of 12 months was 20.2%. The pregnancy rate under in-vivo conditions was significantly higher when semen samples had a better sperm morphology, with significant differences for thresholds at 4, 7 and 14% of strictly normal forms. Although sperm morphology is only one among a multiplicity of factors determining fertility, the results suggest that the evaluation of sperm morphology using strict criteria provides valuable information during basic infertility investigations.

Screening for subclinical inflammation in ejaculates

To determine the clinical significance of albumin determination in ejaculates by means of an easy office test to screen semen samples for subclinical infection-inflammation. One hundred fifty-nine randomly chosen males of couples with longstanding infertility (median duration of infertility 4 years (range 1 to 19 years) without clinical signs or symptoms of genital tract infection. Outpatient Infertility Clinic of the University of Heidelberg, Germany. Screening of ejaculates for subclinical infection-inflammation by means of a ready-to-use kit for semiquantitative detection of albumin in addition to determination of leukocytes rates by means of monoclonal antibodies for differentiation of round cells and measurement of granulocyte elastase concentration in semen samples. Evaluation of sperm quality by means of standard sperm analysis including determination of local antisperm antibodies with the mixed antiglobulin reaction, evaluation of sperm functional capacity in vitro with the standardized sperm-cervical mucus (CM) penetration test, and semen cultures. All tests were performed from aliquots of the same ejaculates. Screening of semen samples for elevated albumin with the modified paper strips proved to be very easy, quick, and suitable for routine use. Positive results were not related markedly to medical history and outcome of clinical examination as well as to standard parameters of sperm analysis and were not influenced by local antisperm antibodies of the immunoglobulin (Ig)G and/or IgA class and microbial colonization. However, albumin-positive semen samples were significantly less frequent in case of very good outcome of the sperm-CM penetration test. A significant relationship was found with high rates of leukocytes of the round cells in semen samples (total range 0% to 96%) and the concentration of granulocyte elastase (total range 1 to 880 micrograms/L). The results of this prospective study suggest that the determination of albumin in semen samples with ready-to-use test kits might be a valuable additional marker for subclinical infection-inflammation of the male genital tract and therefore suitable for screening during infertility investigation.

Anaerobes in ejaculates of subfertile men.

The clinical significance of micro-organisms in semen samples of asymptomatic subfertile patients is a matter of constant debate. Usually little attention is paid to anaerobic bacteria as they are sensitive to transportation and culturing, and differentiation is difficult, costly and time-consuming. In the present study, special screening was carried out for anaerobes in ejaculates in addition to the routine microbial cultures of genital secretions of both partners. In addition to standard semen analysis and evaluation of sperm ability to penetrate cervical mucus (CM) in vivo (post-coital testing) and in vitro using a standardized test system, semen samples from 126 randomly chosen males of couples with a median duration of infertility of 4 years were examined for colonization with anaerobic bacteria. All couples were without clinical signs or symptoms of genital tract infection. The special care taken for anaerobic growth in semen samples gave a high rate of positive cultures and showed that nearly all ejaculates (99%) were colonized with anaerobic micro-organisms, and potentially pathogenic species were found in 71% of men. This rate was more than four times higher than that obtained with routine cultures and standard transportation (16%). Anaerobic bacterial growth of > or = 10(6) colony forming units (CFU)/ml was seen in 42% (total range 10(3)-10(8) CFU/ ml). In addition, aerobic growth was found in 96% (> or = 10(6) CFU/ml in 21%), potentially pathogenic species in 61% of semen specimens. There were no marked differences in the prevalence of anaerobic micro-organisms in patients with reduced or normal sperm count, motility or morphology. Nor was there any significant difference in anaerobic colonization between samples with impaired or good ability to penetrate CM of female partners (in vivo or in vitro), or the CM of fertile donors in the in-vitro sperm-cervical mucus penetration test (SCMPT) in this asymptomatic group of patients. There was no clear association between microbial colonization and subsequent fertility in vivo within an observation period of 6 months. The results of this study suggest that anaerobic bacteria are often not detected when routine methods for microbial evaluation are used. This should be considered during assisted reproduction and in patients with symptoms of genital tract infection and should lead to further studies in infertile patients where subclinical infection or inflammation is indicated by specific markers in semen samples.

Analysis of spermatozoa from the proximal vas deferens of vasectomized men.

This study assessed the condition of spermatozoa from the proximal vas deferens of men after vasectomy. The fluids of both proximal vas deferens were collected from 67 vasectomized men by cannulating the vas deferens at the time of vasectomy reversal. Selected sperm parameters were analysed after incubation of the spermatozoa for 30 min at 37 degrees C. Sperm concentration in the proximal vas from vasectomized men (16,312 +/- 21,496 million per ml, geometric mean: 7948 +/- 398 million per ml) was significantly higher than that of fertile men and was maintained at a constant level independent of the duration of vas obstruction. The means of sperm motility (36.2 +/- 26.2%), spermatozoa with normal morphology (50.7 +/- 21.7%), sperm viability (53.0 +/- 25.3%) and hypo-osmotic swelling test (HOS-test, 53.9 +/- 21.7%) were statistically lower than the respective values for normal fertile men. There was no significant correlation between the duration of vas obstruction and the above semen parameters. In 46.4% of vas fluids all spermatozoa were immotile and this condition was more common after 3 years of vasectomy. Immotile spermatozoa in the proximal vas fluids at the time of vasectomy reversal may be an important factor for predicting semen quality and fertilizing ability after vasovasostomy. There were no significant differences in the results of sperm-cervical mucus penetration test (CMPT) between spermatozoa from vasectomized and fertile men. Antisperm antibodies on the surface of spermatozoa from the vas of vasectomized men were determined by the immunobead test (IBT; 78.6% for IgG, 32.1% for IgA) and sperm cervical mucus contact test (SCMC, 36.4%).(ABSTRACT TRUNCATED AT 250 WORDS)

Immunology: Determination of antisperm antibodies in serum samples by means of enzyme-linked immunosorbent assay—a procedure to be recommended during infertility investigation?*

The clinical significance of antispermatozoal antibody (ASA) testing in serum samples with an enzyme-linked immunosorbent (ELISA) technique using a commercially available kit was evaluated in 95 randomly chosen couples under infertility investigation. Results were related to many other parameters of male and female fertility, e.g. outcome of sperm analysis, testing of sperm functional capacity including the crossed in-vitro sperm-cervical mucus penetration test (SCMPT), results of a microbial screening in genital secretions, testing for local IgG- and IgA-class sperm antibodies in semen by means of the mixed antiglobulin reaction (MAR) and the subsequent pregnancy rate in a prospective study. Results of ASA testing did not show any relationship with medical history and results of clinical examination, the semen quality including sperm function tests and seminal cultures, outcome of the crossed SCMPT, and local ASA. All patients with MAR (IgG or IgA) positive ejaculates were ELISA negative in serum samples. No significant difference with regard to circulating ASA (ELISA) was found in couples with and without a subsequent pregnancy. Furthermore there was no significant difference of antibody levels in subfertile female patients, virgins (n = 36), 'normal' pregnant women (n = 39) and prostitutes (n = 40). In summary, the results of this study clearly demonstrate that the use of this method for detection of antisperm antibodies during infertility investigation cannot be recommended.

The pH as an important determinant of sperm-mucus interaction

To determine the clinical significance of endocervical mucus pH on sperm-mucus interaction during infertility investigation. Two hundred sixteen couples with a median duration of infertility of 4 years (range, 1 to 19 years) presenting at the infertility unit of the Women's University Hospital of Heidelberg, Germany. Determination of endocervical pH by colorimetric and electrometric measurement and correlation of results with the outcome of postcoital testing (PCT) and other parameters of infertility investigation (semen and cervical mucus [CM] quality, microbial colonization of cervix and ejaculates, medical history, hormonal status, and specific medication) and the subsequent fertility in a prospective study. In vitro experiments with the sperm-cervical mucus penetration test (SCMPT) used as biological model. The colorimetric determination of endocervical mucus pH is an easy method, suitable for routine clinical use, correlating significantly with electrometric measurement of pH. Median pH was 7.0 (range, 5.4 to 8.2). The mucus pH was significantly related with the results of PCT, even when mucus and semen variables were taken into account. No significant relationship was seen between the cervical index and mucus pH and the microbial colonization of cervix and ejaculates. The pH of endocervical secretions correlated with the peripheral hormonal status: low pH levels were significantly more frequent in patients with hyperandrogenemia, indicated by high testosterone and/or dehydroepiandrosterone sulfate levels before medication was started, and in hyperandrogenemic patients treated with dexamethasone than in the other women. Oral administration of estrogens led to a subtle alkalinization of the CM. With regard to subsequent fertility 6 months after pH testing, the pregnancy rate was significantly lower in women offering reduced mucus pH on occasion of the PCT in the group of couples with primary infertility and in couples with oligozoospermia of the male partner. The significant influence of pH on sperm-mucus interaction was confirmed in vitro with the SCMPT. The results indicate that the pH of the CM, easily determined with pH indicator paper, is an important parameter of mucus quality with significant influence on spermatozoal viability in CM, which correlates with peripheral hormonal status and can be affected by oral medication with estrogens. Therefore the routine determination of pH on occasion of the PCT is recommended during infertility investigation.

Differentiation of round cells in semen by means of monoclonal antibodies and relationship with male fertility

To differentiate round cells in semen samples of subfertile men and evaluate the clinical significance during infertility investigation. One hundred and eight randomly chosen couples with a median duration of infertility of 4 (range, 1 to 20) years presenting at the outpatient infertility clinic of the University of Heidelberg, Germany. Differentiation of round cells in semen by means of monoclonal antibodies (mABs) and a streptavidin-biotin system for staining. Correlation of results with medical history, outcome of clinical examination, sperm analysis, microbial screening of both partners, evaluation of sperm functional capacity in vivo by means of the postcoital test (PCT) and in vitro with the standardized crossed sperm-cervical mucus penetration test (SCMPT) and the subsequent fertility in a prospective study. The method used for differentiation of round cells proved to be practical and suitable for routine use. The percentage of leukocytes ranged from 0% to 58% with a median of 3%. Number of round cells and percentage of leukocytes did not differ markedly with regard to andrologic history, clinical findings, for example, varicocele, results of standard sperm analysis, and microbial colonization of semen samples. However, high rates of leukocytes of the round cells correlated with reduced sperm count and morphology and results of PCT. Leukocyte-positive (> 15% leukocytes) specimens were also significantly more frequent in case of inadequate SCMPT and reduced sperm penetration ability in vitro. In asymptomatic patients (in terms of genital tract infection), the majority of round cells consist of immature germ cells and < 5% are white blood cells. The streptavidin-biotin system and the mABs used in this study proved to be useful to identify patients with elevated rates of leukocytes in semen possibly reflecting subclinical genital tract infection with influence on sperm functional capacity and subsequent fertility. Thus the procedure can be recommended to be included in a comprehensive evaluation of male fertility.

Dosage frequency and drug-compliance behaviour ? a comparative study on compliance with a medication to be taken twice or four times daily

The objective of the study was to investigate patient compliance with two different dosage regimens. Ethinyloestradiol 80 micrograms daily was prescribed to 35 female outpatients to be taken as 20 micrograms four times daily or 40 micrograms twice daily for 7 days. It was given to standardise cervical mucus before a sperm cervical mucus penetration-test (SCMPT) was performed. Sixty-five patients with primary infertility (mean age 29.9 y) completed the study. Compliance was assessed by microprocessor-based compliance monitoring. Besides compliance (percentage of prescribed doses taken), the adherence of the patients to the dosage schedule was evaluated--'regimen compliance'. The latter parameter of drug intake behaviour was calculated by the number of days in which 2 (BID) or 4 openings (QID) were recorded by the electronic monitor. As a third parameter, the deviation from the prescribed dosing intervals, i.e. 12 or 6 h, was also determined. Partial compliance was the predominant finding and only 9 patients (13.8%) were over-compliant. Mean compliance was 75.7% in all 65 patients as a group, range 7.1 to 143%. The mean compliance with the QID regimen was 67% compared to 85% with the BID regimen. 'Regimen compliance', the percentage of doses taken on schedule, was 36% and 63% for the QID and BID regimens, respectively. Drug-intake behaviour was more erratic with the QID than the BID regimen, as indicated by the 55% of opening intervals recorded which exceeded the range of 3-9 h (mean: 6 h), compared to only 19% exceeding 6-18 h intervals (mean: 12 h).(ABSTRACT TRUNCATED AT 250 WORDS)

Relationship between local anti-sperm antibodies and sperm–mucus interaction in vitro and in vivo*

Sperm-mucus interaction under in-vitro or in-vivo conditions can be affected by local anti-sperm antibodies (ASA). In a prospective study, a mixed antiglobulin reaction (MAR) test, using immunoglobulin-coated red cells and spermatozoa, was used to detect the presence of IgG and/or IgA on the spermatozoal surface in 209 males of infertile partnerships. The results of direct MAR tests range from 0 to 100% and a significant correlation between MAR % IgG and MAR % IgA (r = 0.74) was found. MAR positive semen samples were significantly more frequent in the group of patients with an inadequate outcome of postcoital testing (PCT). The sperm-mucus interaction in vitro, evaluated by means of the in-vitro sperm-cervical mucus penetration test (SCMPT) with cervical mucus (CM) of patients' partners, was particularly related to spermatozoal surface antibodies of the IgA class: MAR IgA positive ejaculates were seen in 13.9% of males with inadequate SCMPT versus 4.4% when SCMPT was adequate. The significant correlation between MAR IgA and the sperm penetration ability in vitro could also be proven when donors' CM was taken for the SCMPT, but not when a non-human material was used for in-vitro penetration testing. Microbial colonization of semen specimens did not interfere with the outcome of MAR testing. The pregnancy rate after 12 months was 23% (48/209) and was significantly lower when greater than 30% of spermatozoa were covered with surface ASA of the IgG or IgA class.(ABSTRACT TRUNCATED AT 250 WORDS)

The use of hens' egg white as a substitute for human cervical mucus in assessing human infertility

Semen samples from 179 patients with longstanding infertility were allowed to penetrate capillaries filled with a fraction of fresh hens' egg white (HEW) as a substitute for cervical mucus (CM) for the in-vitro sperm-cervical mucus penetration test (SCPMT). Results were correlated with those obtained from human CM (HCM) used in parallel on the same semen sample, compared to results of sperm analysis and post-coital testing (PCT) and analysed for their prognostic value for a subsequent pregnancy in a prospective study. The overall pregnancy rate after 6 months was 26.3% (47/179). Whereas routine semen analysis (semen volume, pH, sperm count, progressive motility, morphology and fructose concentration) did not differentiate between patients who achieved pregnancy and those who did not, significant differences were found for the penetration distance (P less than 0.02) and the motility grade of sperm in HEW (P less than 0.02). Although the mean sperm velocity in HEW was reduced compared to HCM, a close positive correlation was found between the sperm density and the penetration distance (P less than 0.001). The percentage of samples with good HEW penetration increased significantly in parallel with better results of post-coital testing (P less than 0.001). The results indicate that, when fresh human CM is not available, HEW can be used as a medium for the SCPMT as a valuable adjunct to semen analysis which provides information about sperm functional capacity.

Clinical significance of crossed in vitro sperm-cervical mucus penetration test in infertility investigation

To evaluate the clinical significance of in vivo and in vitro testing of sperm ability to penetrate cervical mucus (CM), postcoital testing (PCT) and in vitro sperm-cervical mucus penetration testing were compared in a prospective study. Both in vivo and in vitro tests were standardized and performed after an oral course of estrogen therapy. Crossed in vitro sperm-cervical mucus penetration test, evaluated in 277 couples with CM of patients' wives and additionally with CM and semen of fertile donors, revealed that the male factor contributed to a significantly higher extent to deficient sperm-mucus interaction than the cervical factor. The overall pregnancy rate after 6 months was 23% (64/277). Whereas the outcome of PCT did not significantly predict subsequent fertility (PCT good pregnancy rate 24%/PCT poor 20%), significant differences were found for the sperm-cervical mucus penetration test with CM of patients' wives (pregnancy rate, 30.5% versus 8.5%) and for in vitro testing with donors' CM, but not for the mucus penetration test with donors' spermatozoa. Routine sperm analysis did not prove to be of prognostic value for a subsequent pregnancy. The results suggest that the in vitro sperm-cervical mucus penetration test is a good parameter of sperm function and, in particular, when performed as a cross-matching penetrability test, a valuable adjunct to PCT with regard to fertility prognosis.

Prognostic value of in vitro sperm penetration into hormonally standardized human cervical mucus

To analyze the prognostic value of the sperm cervical mucus penetration test (SCMPT), fresh semen samples of 99 male patients under infertility investigation were exposed to capillary tubes filled with freshly obtained cervical mucus (CM) of the patients' wives (WCM), fertile donors (DCM), and bovine CM (BCM). The quality of the human CM was standardized by oral administration of estrogens. The overall pregnancy rate after 6 months was 17.2% (17/99), and was significantly different in couples with poor and good SCMPT with WCM (1/44, 2.3% versus 16/55, 29%; P less than 0.001) in a prospective study. Human CM was superior to BCM as a penetration medium in providing more information about sperm function. The results suggest that in vitro sperm penetration testing with hormonally standardized CM of female partners adds an important dimension to sperm analysis with regard to fertility prognosis.

Results of in vitro fertilization in women with antisperm antibodies in serum, cervical mucus, and follicular fluid.

This study was undertaken to investigate the presence of antisperm antibodies (ASA) in serum, cervical mucus, and follicular fluid (FF) of women undergoing in vitro fertilization and embryo transfer (IVF-ET). IgG and IgA ASA directed mostly against sperm head were found at similar concentrations in serum, cervical mucus, and FF of 2 of 34 patients. Ninety-one percent fertilization and 100% cleavage rates, respectively, were observed in one of the two patients. No fertilization occurred in the second patient. In both women, in vitro sperm penetration tests revealed hostile mucus and repeated postcoital tests were poor. It is concluded that the sperm-cervical mucus penetration test and mucus ASA measurements are useful in establishing the diagnosis of immunological infertility.

Correlation of postcoital evaluation with in vitro sperm cervical mucus determinations and Ureaplasma cultures

Fifteen trials were completed in 14 couples during an infertility evaluation. The postcoital test (PCT) was accomplished in a standardized manner. Also, standardized determinations of the sperm-cervical mucus penetration test (SCMPT) with the addition of cross-testing (X-test) utilizing bovine cervical mucus (BCM) and normal donor semen (NDS) were done. Male and female samples were also cultured for Ureaplasma urealyticum using U9-B indicator broth and A-7 agar. The PCT and SCMPT agreed in 87% (13/15) of the cases. Utilizing BCM and NDS, where possible, the causative factor was the cervical mucus in 54% (7/13); semen factor in 15% (2/13); both factors in 8% (1/13); and undetermined in 23% (3/13). U. urealyticum cultures were positive in 40% (6/15) of the cases. Analysis of the results revealed: (1) significant correlation between PCT and SCMPT (P less than 0.01); (2) no significant difference among the PCT, SCMPT, and X-test, indicating that the cervical mucus was the causative factor; and (3) no U. urealyticum correlation with the PCT or the SCMPT. Thus, laboratory SCMPT and X-test correlated with the PCT, providing additional information concerning the causative factor in infertility. The U. urealyticum status in cervical mucus and semen cannot be determined from the PCT nor the SCMPT.