FIELD: medical diagnostics.
SUBSTANCE: perform sampling and processing of biological material; after that, carry out diagnostics and, on the basis of the received data make a conclusion about male reproductive function, use ejaculate as biological material. Processing of the ejaculate is carried out as follows: placed an aliquot of 100 mcL of fresh ejaculate on a dry disposable glass slide; place the glass slide in a thermostat at 37°C for 25-30 minutes until it is completely dry; then take out the slide with dried aliquot and left for 2 mins at room temperature 22°С; place the slide with dried aliquot in a sealed plastic container for storage at -22°C for a period of 28 days for subsequent delayed study. After 28 days of storage, before assessment of spermatozoa DNA fragmentation take out the glass slide with a frozen air-dried aliquot, leave it at a temperature of +22°C; add to the dried aliquot of the ejaculate 50 mcL of phosphate-buffered saline; mix with a pipette tip until a homogeneous suspension is formed; place the resulting suspension in 0.5 ml Eppendorf conical microtube; then coat the glass slide with a positively electrically charged surface to ensure that the cellular material is attached to the surface; mix the ejaculate suspension with inert agarose microgel; cool and expose to denaturant and lysing solution; apply to a glass slide; stain and evaluate spermatozoa DNA fragmentation by counting 300 spermatozoa per sample on a laboratory counter using bright field microscopy under x1000 immersion lens; estimate DFI ejaculate fertility by taking into account spermatozoa DNA fragmentation according to the formula. If DFI is less than or equal to 15%, make a conclusion about normal reproductive function, if DFI is greater than 15%, but less than 25%, make a conclusion about doubtful reproductive function, if DFI is greater than 25%, make a conclusion about abnormality.
EFFECT: invention provides increase in the diagnostic reliability of male reproductive function.
3 cl, 1 dwg, 6 tbl
