Identification ofLeukoctyospermia

A wide variety of detection methods are available to identify leukocytes in seminal plasma. Determining the quantity of seminal ROS generation in a sample has been suggested as an important test to be performed in the clinical andrology assessment [57] . The identification of the presence of ROS-producing cells in a semen sample can be an important step taken in establishing the underlying cause for acquired or inherent defects that may be undetected [57] .

The tests carried out to establish the condition of leukocytospermia include immunocytology, peroxidase, polymorphonuclear-elastase, and cytology [7] . The traditional method and technique recommended by the WHO [ 18] for counting leukocytes in human semen is to use a histochemical procedure to identify the per-oxidase enzyme found in the cytoplasm that characterizes PMN-granulocytes [37] . In the examination of a semen sample, the difference between immature germ cells and leukocytes cannot be morphologically distinguished from each other, therefore cytological identification is an inaccurate method to detect leukocytes [68] . PMN-granulocytes and leukocytes are considered "round cells" of nonspermatogenic origin [69]. Due to a lack of diligence in differentiating between different categories of round cells, particularly spermatogenic germ cells and leukocytes, an overestima-tion of the number of leukocytes in a semen sample can often result [70] which can increase the chance of misdiagnoses. There is a significant relationship between the presence of PMN granulocyte elastase and the concentration of peroxidase-positive cells in a semen sample [68]. This particular cytological staining technique is considered to be a reliable method in the detection of leukocytes as it has minimal chances of misdiagnoses [27].

However, it must be considered that during the inflammatory process, degranulation and deregulation occur, resulting in the extracellular liberation of the leukocyte's cellular contents. The quantification and identification of leukocytes based on this specific method may therefore be affected as the peroxidase compound may be undetectable [71].In order to ensure the most accurate quantification of leukocytes, an obvious recommendation would be to use peroxidase staining, which detects intracellular enzymatic activity [67], as well as an alternative method such as the biochemical analysis of the concentration of PMN-elastase, which identifies extracellular enzymes.

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