Infertility Diagnostics

Last updated: 10 September 2025

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Laboratory Tests and Ancillaries

Diagnostic Assessment  

Most couples will require the following diagnostic tests:

Semen Analysis



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Semen analysis is done to exclude abnormalities in the semen and to rule out azoospermia. Confirmatory tests are ideally repeated after 3 months or as soon as possible if there is gross deficiency.

World Health Organization's (WHO) Accepted Reference Values for Semen Analysis, 2021

Parameter (Units)

Reference Value (Lower Limits, Fifth Percentile)

Serum volume (mL)

1.4

pH

≥7.2 

Sperm concentration (106/mL)

16

Total sperm number (106/ejaculate)

39 

Total motility (%)

42

Progressive motility (PR, %)

30

Vitality (live spermatozoa, %)

54

Sperm agglutination

Absent

Sperm morphology (normal forms, %)

WHO criteria: Lower reference limit for normal forms is 4%
Tygerberg strict criteria: Excellent prognosis (>14% morphologically normal spermatozoa), good prognosis (4-14%), poor prognosis (<4%)

References: 1) The Unexplained Fertility Guideline Group, Romualdi D, Ata B, Bhattacharya S, et al. Evidence-based guideline: Unexplained infertility. ESHRE. https://www.eshre.eu/. 2023. 2) World Health Organization (WHO). WHO laboratory manual for the examination and processing of human semen. 6th ed. WHO. https://www.who.int/. 2021. 

Assessment of Ovulatory Function  

Ovarian reserve, which is the number of oocytes available for fertilization at a point in time, may be assessed through serum tests or ultrasonography. Tests include cycle day three follicle-stimulating hormone or estradiol, Clomifene citrate challenge test, anti-Müllerian hormone (AMH), or antral follicle count (AFC).  AFC and AMH are the most simple, sensitive, and specific tests for ovarian reserve. The presence of reduced ovarian reserve predicts future response to ovarian stimulation. Tests to determine ovarian reserve are not necessary to identify the etiology of infertility or to predict the probability of spontaneous conception over 6-12 months in women with regular menstrual cycles.  

Inquire about the frequency and regularity of menstrual cycles. Women with regular cycles are likely to be ovulating. In women with regular menstrual cycles, perform a midluteal phase serum progesterone level (day 21 of a 28-day cycle) to confirm ovulation. The European Society of Human Reproduction and Embryology (ESHRE) does not routinely recommend tests to confirm ovulation in women with regular menstrual cycles but if needed, tests (eg urinary luteinizing hormone measurement, ultrasound [US] monitoring, mid-luteal progesterone measurement) may be performed to confirm ovulation. In women with prolonged irregular cycles, serum gonadotropins (follicle-stimulating hormone and luteinizing hormone) and serum progesterone should be measured. Serum progesterone should be measured based on timing of menstrual periods, which may be done later in the cycle and then repeated every week until the next menstrual cycle starts. Prolactin and thyroid function should only be evaluated in patients with signs and symptoms of thyroid dysfunction, ovulatory disorder, galactorrhea, or pituitary tumor. Testosterone level may also be checked for hyperandrogenism.  

Other Diagnostic Tests  

Initial laboratory tests include complete blood count (CBC), urinalysis, screening for sexually transmitted infections (eg Chlamydia trachomatis), rubella and varicella titers, and Pap smear. Screening for other endocrine diseases is not routine and is done only based on findings obtained from history and physical examination. Monitoring and charting basal body temperature is unreliable and is no longer recommended. Post-coital testing of cervical mucus has no predictive value and is also not recommended in couples with unexplained infertility.

Imaging

Further Evaluation  

Assessment of Tubal Patency  

Prior to instrumental testing of tubal patency, women should be offered a screening test for Chlamydia trachomatis.  

Hysterosalpingography (HSG)/Sonohysterogram



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Sonohysterogram is usually preferred in women without known comorbidities (eg pelvic inflammatory disease [PID], previous ectopic pregnancy, endometriosis) to rule out tubal occlusion.  

Laparoscopy  

Laparoscopy may be preferred in women with known comorbidities so that tubal and other pelvic pathologies can be investigated. This is considered when hysterosalpingography suggests tubal disease that may be repaired. A routine diagnostic laparoscopy is not recommended to diagnose unexplained infertility.  

Assessment of Uterine Cavity  

Two-dimensional (2D) or Three-dimensional (3D) Transvaginal Ultrasound (US)  

Two-dimensional (2D) or three-dimensional (3D) transvaginal ultrasound helps in the detection of uterine leiomyomas affecting the uterine cavity. The use of 3D ultrasound improves the detection of Müllerian anomalies, and its diagnostic accuracy is comparable with a magnetic resonance imaging (MRI). A 3D ultrasound is preferred to rule out uterine anomalies in women with unexplained infertility. This may be used to distinguish between a uterine septum and a bicornuate uterus.

Sonohysterography  

Saline infusion sonohysterography provides information about the endometrial cavity, myometrium and adnexa and is the preferred imaging modality to assess the uterine cavity. This is better than a routine ultrasound in terms of diagnosing intrauterine adhesions, polyps and congenital uterine anomalies.  

Hysteroscopy



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Hysteroscopy is a definitive modality for the evaluation of endometrial cavity abnormalities. This is indicated for the confirmation and treatment of intracavitary lesions detected by other imaging modalities. An advantage of hysteroscopy is that it offers an opportunity for treatment at the time of diagnosis when performed in the operating room. This is not routinely used for initial evaluation of women with infertility due to cost and access considerations.  

Hysterosalpingography  

Hysterosalpingography can detect developmental or acquired anomalies of the uterine cavity which can negatively impact fertility, including submucous fibroids, polyps, synechia, a T-shaped cavity, and congenital Müllerian anomalies. Abnormalities identified on hysterosalpingography require further evaluation with other imaging modalities such as 3D ultrasound, hysteroscopy, MRI or laparoscopy.