Aniline blue staining as a marker of sperm chromatin defects associated with different semen characteristics discriminates between proven fertile and suspected infertile men

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Abstract
Summary: A retrospective study of 49 men with proven fertility and 396 suspected infertile men was conducted with the primary objective of investigating the relationship between the nuclear maturity of sperm and male fertility. Acidic aniline blue staining was used to detect chromatin defects of sperm nuclei related to their nucleoprotein content as associated with DNA. The discriminant value of the percentage of unstained nuclei (= percentage of mature heads, MH) and of other semen characteristics, was analysed by a stepwise (forward) linear regression model. Semen characteristics that discriminated significantly between the two groups of subjects were, in descending order: (1) the percentage of normal sperm, (2) the percentage of amorphous heads, (3) the percentage of tapered heads, (4) semen volume, and (5) the percentage of MH. The discriminant value of each of the significant characteristics was studied by means of ROC‐curves. MH had the best ROC‐curve profile; its cut‐off value was found to be equal to 70% (74.5 ± 2.6% for the donor group versus 53.0 ± 1.1% for the patient group). A simple infertility score (SIS) including MH, was built according to the cut‐off values inferred from the ROC‐curves. SIS allowed an overall satisfactory separation of the two groups (≤4 = fertile, 5–6 = uncertainty zone, >6 = infertile). Our results indicate that the addition of the evaluation of sperm head maturity to routine semen analysis improves the assessment of fertility in men.