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- ArticlePragay DA, Casey SJ, Gotthelf J.Clin Biochem. 1977 Oct;10(5):183-7.Generally acid phosphatase (ACP) assay is used for testing cases of alleged rape. Comparison of three different chemical methods for Acid Phosphatase (Andersch's method with p-nitrophenyl-phosphate substrate and tartrate inhibitor (A-Tart), Roy's method with thymolphthalein phosphate substrate (R-TMP), and Babson's method with alphanaphthyl phosphate substrate) indicated that Roy's method (R-TMP) should be the preferred one. This method had both acceptable sensitivity and confirmed specificity. Our data confirmed that the vaginal wash of normal healthy women has a very low level of ACP activity. Because of inconclusive data in the literature regarding this ACP level, a normal and equivocal range of ACP was suggested until more is known about causes and interferences. Possible sources of normal ACP activity in the wash fluids were also indicated.