The andrological examination allows to highlight through the inspection and palpation of the male genitals, the possible presence of anatomical abnormalities and/or alterations caused by infectious or traumatic processes.
EXAMINATION OF THE SEMINAL FLUID (SPERMIOGRAM)
The standard seminal fluid examination or spermiogram is the starting point for the andrological diagnostic framework. A period of abstinence from sexual intercourse (ejaculation) of between 3 and 5 days (not exceeding) is required to perform the examination.
It is essential to wash your hands and genitals thoroughly before collecting in the sterile urine container.
The examination consists of the evaluation of macroscopic and microscopic parameters of the ejaculate and is carried out in the laboratory in compliance with international standards proposed by the World Health Organization (WHO 2010).
The evaluation of macroscopic parameters includes: Fluidification, Viscosity, Appearance, Volume and pH.
The evaluation of the microscopic component concerns the gametic fraction (spermatozoa) and the non gametic fraction (round cells, epithelial cells), therefore it is analyzed:
– Sperm concentration per unit volume.
– Percentage of mobile forms
– Concentration of mobile forms per unit volume
– Morphology of the spermatozoa present in the sample
– Round Cells
– Blood cells
This examination is not able to determine male fertility in an absolute way because sperm production by the testicles is subject to high intraindividual variability. Therefore, proven paternity is the only proven proof of fertility.
NEMASPERMIC SEPARATION TEST
This method allows you to select the “best” fraction of sperm for motility and morphology. There are several procedures to obtain this selection (Figure 1):
- Swim-up (ascending migration)
- Density gradient separation
The choice of the metology to be used depends on the characteristics of the seminal fluid to be treated: in the case of normozoospermic samples the Swim-up technique is used; in cases of severe oligozoospermia or astenozoospermia it is preferable to adopt the separation on density gradient. Another important difference between these two methods is represented by the different degree of contaminants eventually present in the seminal plasma. The seminal fluid may also contain infectious agents, in which case the technique to be used is preferably the density gradient followed by Swim-up in order to remove the infectious agent as far as possible. However, it must be considered that both procedures cannot guarantee 100% elimination of the potential infectious agent.
SEARCH FOR ANTISPERM AUTOANTIBODIES
Antisperm antibodies are proteins that bind to the sperm surface and inhibit their functionality, interfering with the fertilization process. This test is carried out on the seminal fluid sample to assess the presence of two main classes of immunoglobulins: IgA and IgG.
SPERM DNA FRAGMENTATION TEST
The DNA fragmentation test is performed on the seminal fluid sample to evaluate any structural alterations in the genetic material of the spermatozoa. The evaluation of the percentage of spermatozoa possessing fragmented DNA is carried out through the use of enzymes, able to identify the presence of breaking points present in the DNA double helix. The method used is TUNEL (acronym of Terminal deoxynucleotidyl transferase dUTP Nick End Labeling). Seminal fluids in which the percentage of sperm with fragmented DNA exceeds 30% are considered pathological (Chohan et al 2006).
F.I.S.H. (Fluorescence In-Situ Hybridization) is a genetic analysis that is performed on the seminal fluid sample to assess the possible presence of numerical abnormalities of chromosomes (generally 13, 18, 21, X and Y) in spermatooza. This method involves the use of marked probes that bind specifically to the DNA of the spermatozoa and can be visualized by fluorescence microscopy. If two pairs are present for each chromosome analysed, this is called diploidy. In patients with serious seminal fluid pathologies there is a higher frequency of numerical alterations of the sex chromosomes X and Y that can be inherited from the progeny.
The vitality test is carried out in cases where a marked hypokinesis is observed, to define whether the condition is caused by defects in the spermatozoon’s motor apparatus or by cell death. It is carried out by means of an eosin test using a vital dye that distinguishes living cells from dead cells.
It is carried out on a sample of seminal fluid to investigate the possible presence of microbiological populations that can cause inflammatory processes in the genital tract and sometimes compromise reproductive function.
MALE SCREENING FOR IVF TREATMENT
Preliminary laboratory testing package for IVF
– Blood type and Rh factor
– HB electrophoresis
– HbsAg, HBcAb IgG-IgM, HCV, HIV, TPHA and VDRL (90 days of validity)*.
– CMV (IgG, IgM)
– Preconceptional Genetic Analysis (Most Frequent Hereditary Diseases)
*further examinations of infectious diseases in case of specific indication (e.g. malaria, HTLV-I, etc.).