What Tests Should Be Done to Diagnose Infertility in Men?

Abstract

This study evaluated male infertility through a semen analysis and blood tests. It accounts for 40-50% of the infertility rate and affects 07% of all men. There are different ways to diagnose male infertility which help for the proper diagnosis of the disease and its causes and treatment also some diagnoses are as follows – semen analysis, assessing sperm concentration, motility, morphology, measuring hormones like FSH, LH, Testosterone, and prolactin for endocrine function and other advance tests. After the assessment of the tests the identifying potential causes of male infertility for targeted treatment options, including lifestyle changes and modifications, medication, or assisted reproductive technologies.

Introduction

Male infertility is the inability to cause pregnancy in a fertile female. There are different causes for male infertility hypospermia, Oligoasthenoteratozoospermia (OAT), alkaline PH due to Infections, liquification disorders, Anabolic steroids (steroid activity like growth), glandular secretory activity/enzymes, Peculiar odor.

Diagnostic points can be relatable by history of the patient like Alcohol, tobacco, drugs abuse, liver and kidney diseases (can cause male infertility by disrupting hormone balance, impacting sperm production, and causing erectile dysfunction), ejaculatory dysfunction, psychiatric drugs (selective serotonin reuptake inhibitors-SSRIs and tricyclic antidepressants TCAs), respiratory diseases (Kartagener syndrome/ young’s syndrome), infection like lumps, obesity, sexual dysfunctions. physical examination (Klinefelter syndrome, pituitary adenoma, abnormal curvature/Peyronie’s, undescended testicle, orchitis/consistency/size, epididymitis/cyst, presence of varicocele, size, and consistency), semen analysis (sperm count, motility, morphology), hormone blood tests, scrotal ultrasound to check for varicoceles, and in some cases, a testicular biopsy to evaluate sperm production if semen analysis shows very low or no sperm count.

Tests to be performed for the diagnosis of male infertility are as follows –

1. Semen Analysis Test

which includes the following seminal parameters –

• Semen Volume – The amount of semen in an ejaculation (ml) >= 1.5 mL. If the count is less than 1.5 ml, the condition is called Aspermia.
• Sperm Count – The number of sperm per milliliter of semen > 39 million. If the values are low then the condition is called oligospermia or azoospermia.
• Sperm total motility (PR – NP) – PR stands for Progressive Motility, meaning sperm that are actively swimming in a forward, straight line, while NP stands for Non-Progressive Motility, indicating sperm that are moving but not traveling in a straight line, essentially swimming in circles or with poor forward progression together these categories make up the total sperm motility which ranges as >40%(38-42). If the range is less than 38 it is called teratozoospermia.
• Progressive motility means sperm that are actively swimming in a forward, straight line (PR,%) is >32% (31-34)
• Vitality (Live spermatozoa, in % means male sex cells that are alive and can fertilize a female egg) – >58% (55-63)
• Sperm morphology (Normal forms, %) >4 % (3.0-4.0)
• pH value means acidity or alkalinity of the semen having value > 7.2
• Peroxidase-positive leukocytes mean the presence of white blood cells in semen this is called leukocytopenia ranges
• Viscosity

How to prepare

A person should avoid ejaculation for 24 – 72 hours before the test, avoid alcohol, caffeine, and drugs like cocaine and marijuana for 2 – 5 days before the test, and stop taking herbal medicine like Echinacea (Kalmegh) and hormone medication for proper results.

How to collect a sample

Masturbate in a sterile jar or cup or a special condom and submit the sample in the lab before 30 min.

2. Optional Investigations Are

• MAR test (Mixed antiglobulin reaction test) should be
• Immunobead test (motile spermatozoa with bound beads)
• Seminal Zinc (for prostate function) is > 2.4 mmol/ejaculate
• Seminal fructose (a marker for seminal vesicle function) is > 13 mmol/ejaculate
• Seminal neutral glucosidase is

How to prepare

A person should avoid ejaculation for 24 – 72 hours before the test, avoid, alcohol, caffeine, and drugs like cocaine and marijuana for 2 – 5 days before the test, and stop taking herbal medicine like Echinacea (Kalmegh) and hormone medication for proper results.

How to collect a sample

Masturbate in a sterile jar or cup or a special condom and submit the sample in the lab before 30 min.

3. Semen Culture

Semen culture is also called sperm culture This is a laboratory test done to check for bacteria in semen. It is often performed to evaluate male fertility. It includes some points in the results as follows –

• Antimicrobial Susceptibility Testing (AST) – A laboratory procedure that determines how bacteria respond to antimicrobial agents.
• Organism Identification – If it’s positive it shows the presence of the specific type of bacteria in the semen sample.
• Pathogen Detection – The test checks for the presence of common pathogenic microorganisms, such as mycoplasma and ureaplasma.

4. Leukocytospermia

A condition with more than 106 WBC per milliliter of semen cells present in semen. It can be a sign of infection or inflammation, which can negatively impact on semen quality and fertilization. Tests to detect WBCs in semen are –

Peroxidase Test

A test that quantifies granulocytes, the most common WBC type in semen

5. Immunocytology

A technique that uses antibodies to identify and analyze proteins in cells. It combines cytology (the study of cells) and immunology (the study of the immune system).

How to perform

Immunohistology involves examining smears with antibodies. The antibodies may include pan-cytokeratin, human epithelial antigen 125, and calretinin. Cells may need to be permeabilized with detergents like Triton X-100 or Tween-20. Cells may need to be fixed with organic fixatives like acetone, methanol, and ethanol.

Uses Of Immunocytology Test

Immunocytology is used to diagnose, monitor, and predict diseases. It’s used to identify abnormal cellular behavior, such as in malignant and benign disorders. It’s used to detect metastatic carcinoma, neuroblastoma, and bladder cancer.

Leukocyte Esterase Dipstick Test

Test that detects WBC in semen. a rapid urine test that detects the presence of an enzyme called leukocyte esterase, which is released by white blood cells, essentially acting as a screening tool to indicate a possible urinary tract infection (UTI) by identifying elevated levels of white blood cells in the urine. Leukocytospermia can be assessed using methods like the Endtz test, the peroxidase test, and flow cytometry.

Flow Cytometry

A test that uses light scatter properties to identify WBC populations or particles in a fluid.

How To Perform

Cells are stained with a dye to make them more sensitive to light. The cells are placed in a fluid and passed through a narrow stream. A laser excites the cells, causing them to emit light. The light is detected by a system of filters, mirrors, and detectors. The light signals are converted into electronic signals and analyzed by a computer.

Endtz Test Also Called Myeloperoxidase Test

It’s a type of semen analysis that identifies the presence of WBCs and immature sperm and is used to identify peroxidase-positive granulocytes, such as neutrophils and macrophages.

What it indicates

A positive Endtz test result indicates the presence of excessive reactive oxygen species (ROS) in the semen and a high WBC count in semen is often associated with male infertility.

A Hormonal Test To Check Fertility In Males

Typically involves a blood test to measure levels of hormones like testosterone, follicle-stimulating hormone (FSH), and luteinizing hormone (LH).

Low sperm count and hormones relation – A person with a sperm count

1. Condition when there is low testosterone level, sperm count is low, high LH and FSH levels – This indicates primary hypogonadotropic hypogonadism, which means both spermatogenesis and Leydig cell function are affected. Through karyotyping, we can check chromosomal anomalies, such as Klinefelter’s syndrome and Y chromosome microdeletion (sperm count is
2. A condition with low testosterone level, low sperm count, and high FSH level is high with normal LH levels – This indicates seminiferous tubule damage and no damage to Leydig cells and abnormal spermatogenesis.
3. When there is low testosterone, low sperm count with normal or low FSH and LH levels, and hypogonadism – Here it is important to check other pituitary hormone deficiencies, which include thyroid function (free T4), prolactin levels, cortisol in the morning. High prolactin levels mean that there can be a prolactin-secreting tumor.
4. If a person has a high testosterone level, low sperm count, and normal FSH level, high level of LH – A person with this condition can have gynecomastia, due to improper androgen receptor function. This may indicate partial androgen insensitivity syndrome in men.
5. If the testosterone level is normal, low sperm count, normal FSH and LH, and normal testis size – It can indicate an obstruction in the genital tract.
6. Estradiol test – It measures the primary type of estrogen in the blood, known as estradiol (E2).
7. Serum progesterone test – Adrenal glands that may cause high levels of progesterone in both females and males. A progesterone level in a male indicates the amount of this hormone present in his body, which plays a role in sperm production and can be a potential marker for certain health conditions, including prostate health when levels are abnormally high.

How to perform

A blood sample is taken from the veins and used for measuring hormone levels like testosterone, LH, FSH, and prolactin.

Genetic testing for male infertility

It involves a blood test to analyze the chromosomes, particularly the Y chromosome to detect the missing genetic information that could affect sperm production these include –

• Cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation testing – This is often done when a man is experiencing low sperm count or azoospermia (no sperm) and can help identify potential genetic causes of infertility and causes infertility in males by affecting the development of the vas deferens.
• Karyotype analysis – Examines the overall number and structure of chromosomes in a blood sample, including the sex chromosomes (X and Y).M
• Y-chromosome microdeletion testing – Specifically looks for deletions on the Y chromosome, a region crucial for sperm production.

Imaging Test

• Ultrasound – A non-invasive test that uses sound waves to create images of the prostate gland and surrounding tissues. Used to detect epididymis cyst, Testicular atrophy, etc.
• MRI – A non-invasive test that can help diagnose male infertility.
• Venogram – An invasive test that can help diagnose male infertility.
• Doppler ultrasound – This specialized ultrasound technique can assess blood flow within the scrotum, which is important for evaluating varicoceles.

Testicular Biopsy

A small amount of testicular tissue is removed to determine how well the testicles produce sperm.

Conclusion

Male infertility is a common problem these days. In a study of male infertility, the causes and diagnosis are based on a comprehensive evaluation of male infertility typically begins with semen analysis as a primary diagnostic tool assessing sperm count, mortality, morphology, and further testing like hormone testing, genetic screening, imaging study (CT, MRI or ultrasound), and other approaches also required. This will help us to rule out the cause of infertility, give clarity in the diagnosis of the main cause and also help in treatment.