Oya M, Kido A
Z Rechtsmed. 1984;91(4):269-78. doi: 10.1007/BF02332320.
A new method for identification of seminal stains is described, based on the immunologic demonstration of leucine aminopeptidase (LAP), which is extremely abundant in human semen and specific for the prostate as well as semen. An antiserum against human seminal plasma was obtained by repeated immunization of rabbits with seminal plasma and Freund's adjuvant. Ouchterlony's double immunodiffusion test and Culliford's precipitin electrophoresis were performed to demonstrate specific proteins of seminal plasma. LAP activity was visualized with L-leucyl-beta-naphthylamide as substrate and with Fast Garnet GBC as coupler. The immunologic analysis of LAP produced two precipitin lines with enzyme activity. One was observed in kidney, jejunum, pancreas, prostate, as well as in semen, and was completely absorbed with kidney homogenates. The other was found only in semen and the prostate and was not absorbed with kidney homogenates. When the anti-seminal plasma serum absorbed with the kidney was used, the semen-specific LAP could be demonstrated by precipitin electrophoresis only in seminal stains stored for up to 2 months, whereas it was not demonstrated in stains from other human body fluids. By means of precipitin electrophoresis the detection of the semen-specific LAP was possible at semen dilutions of up to 1:32. The method described here greatly enhances the value of semen identification and is quite recommendable for the examination of stains in medico-legal practice.
本文描述了一种鉴定精液斑的新方法，该方法基于亮氨酸氨肽酶（LAP）的免疫检测，LAP在人类精液中含量极高，且对前列腺和精液具有特异性。通过用精浆和弗氏佐剂反复免疫兔子获得了抗人精浆的抗血清。采用双向免疫扩散试验（Ouchterlony试验）和沉淀素电泳（Culliford沉淀素电泳）来检测精浆中的特异性蛋白质。以L-亮氨酰-β-萘酰胺为底物、固红GBC为偶联剂来显示LAP活性。对LAP的免疫学分析产生了两条具有酶活性的沉淀线。一条出现在肾脏、空肠、胰腺、前列腺以及精液中，可被肾脏匀浆完全吸收。另一条仅在精液和前列腺中发现，不能被肾脏匀浆吸收。当使用经肾脏吸收的抗精浆血清时，精液特异性LAP仅在保存长达2个月的精液斑中通过沉淀素电泳得以显示，而在其他人体体液斑中则未显示。通过沉淀素电泳，在精液稀释至1:32时仍可检测到精液特异性LAP。本文所述方法极大地提高了精液鉴定的价值，非常推荐用于法医实践中的斑迹检验。
