Normal sperm morphology and chromatin packaging: comparison between aniline blue and chromomycin A3 staining

The successful implementation of ICSI has provided a unique means of allowi ng couples suffering from severe male infertility to achieve their reproduc tive goals. However, despite the great therapeutic advantages of the techni que, ICSI often provides solutions to clinicians in the absence of an aetio logical or pathophysiological diagnosis. The development of a sequential di agnostic schedule for patients consulting for fertility disturbances would be an ideal method of approach. Since sperm morphology recorded by strict c riteria has often been correlated with fertilization failure, the present s tudy aimed to evaluate the relationship between normal morphology and chrom atin staining among fertile and subfertile men. Both chromomycin A3 (CMA3) and acidic aniline blue (AAB) were employed to record chromatin packaging q uality among 58 men visiting the andrology laboratory. Intra- and interassa y variations were initially recorded for fertile sperm donors. The coeffici ents of variation (CV) for all intra- and inter-assay assessments were <12% . Chromatin packaging was significantly and negatively correlated with norm al sperm morphology, namely r=0.40 (P=0.001) and r=0.33 (P=0.001) for CMA3 and AAB, respectively. Receiver operator characteristics illustrated sensit ivity and specificity values of 75% and 82% for CMA3 and 60% and 91% for AA B, respectively. Significantly different CMA3 and AAB staining was recorded among men with severe teratozoospermia (<4% normal forms) when compared wi th normozoospermic men (>14% normal forms)? namely 49% vs. 29% for CMA3 and 51% vs. 26% for AAB staining, respectively. Chromatin packaging assessment s should be a valuable addition to the sequential diagnostic programme in a n assisted reproduction arena.