In forensic most of the rape cases we encounter cloths of victim, offender and other articles like bedsheets, pillow, sofa, and other objects which may have seminal stains to examine these stains we do acid phosphatase for the preliminary examination of these stains and after that microscopic examination for the confirmation of spermatozoa and further DNA can be extracted from these stains. The evidence submitting for examination in the laboratory has been washed intentionally which resulting in washing of semen from the items of examination. To know the persistence of spermatozoa and acid phosphatase activity 3 different types of fabrics has been repeated wash with normal water and detergent water, acid phosphatase test has been performed on them and microscopy of stains has observed in this experiment.