Polymorphism of EsD by isoelectric focusing: phenotyping in human tissues, dental pulps, hair roots, and semen

The polymorphism of EsD was investigated in tissues of various human organs, dental pulps, hair roots, and seminal stains by isoelectric focusing. The method yielded an excellent resolution of the isoenzyme components. The time limits of determination were: in organ tissues 3 weeks, in dental pulps 1 week, and in hair roots several days. The 7-1 type was less stable than the common types. Phenotyping was possible from fresh semen samples, but was unsuccessful from dried seminal stains after storage. The results show that the EsD typing by isoelectric focusing is of practical use for medicolegal individualization of organs, teeth, and hairs.     

用ELISA检测精液及精斑中精子抗原的研究

本实验应用单克隆抗人精子抗体和酶标记羊抗人精子抗体,采用ELISA方法确定精子抗原成份的存在。对10份新鲜精液,15份精斑进行了验测,其结果阳性率为100%。新鲜精液(精子数约10,000万个/ml)稀释100万倍,精斑浸出液稀释50万倍,均可出现阳性。对唾液斑、尿斑、乳汁斑、阴道斑、汗斑及输精管结扎的精液均为阴性。实验结果表明,本法检验精子抗原具有灵敏度高,特异性好的优点。     

The establishment of the presence of sperm in stains by an agar gel electrophoretic method]

Comparative investigation of seminal stains and other secretions from human body and blood as well as objects of plant origin by gel electrophoresis using alkaline values of pH buffer solutions and subsequent enzymography for acid phosphatase made it possible to develop optimal conditions for detection of seminal presence in stains. Good preservation of seminal acid phosphatase in stains during several years is shown.     

alpha-L-fucosidase phenotyping in human placentae, semen and seminal stains

The polymorphism of alpha-L-fucosidase (Fu) was investigated in a Japanese population from samples of placentae and semen, using isoelectric focusing. The gene frequencies of placental types were Fu1 = 0.748 and Fu2 = 0.252, and those of seminal types were Fu1 = 0.739 and Fu2 = 0.261. The coincidence in the distribution between the placental and seminal types suggests that the Fu types occurring in placentae and in semen are controlled by the same Fu alleles. The Fu typing was possible in seminal stains stored at 4 degrees C for up to 9 weeks, at room temperature for up to 7 weeks and at 37 degrees C for up to 4 weeks. The Fu types were still detectable at semen dilutions of up to 1:4. This polymorphism would provide a useful genetic marker for the medicolegal grouping of seminal stains.     

用酶标抗人精液单克隆抗体A_(10)C_6建立ELISA斑点法快速鉴定人精(液)斑

作者用过氧化物酶标记自己研制的抗人精液单克隆抗体A10C6建立了简便快速鉴定人精斑的ELISA斑点法，直接通过酶标抗体上的酶催化底物显示颜色变化，来判断结果，阳性呈灰色斑点，阴性为无色。其结果只对人精斑浸出液显示阳性斑点，对人的其他组织器官及体液均呈阴性，对动物精斑也无交叉反应，精斑浸出液稀释到1：3200倍仍呈阳性结果。     

The use of scanning electron microscopy in the examination of seminal stains

Samples of human seminal stains on fabrics, paper and wood were observed with a scanning electron microscope (S.E.M). It seems a reliable, rapid and simple technique for the routine examination of seminal stains.     

Immunologic characterization of human seminal leucine aminopeptidase (LAP) and its medicolegal use

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.     

The LAP test. A new method for identification of seminal stains by a qualitative color reaction of leucine aminopeptidase.

A new simple method for identification of seminal stains is described. It employs a qualitative color reaction based on histochemical technique for demonstration of leucine aminopeptidase (LAP), which is extremely abundant in human semen. The method herein reported (the LAP test) is quite suitable for medicolegal examination of seminal stains as a preliminary test.     

混合斑中精斑ABO血型的检验

<正> 1988年,壹岐裕志等报告了用吸附抗α_2-SGP 血清的硝化纤维素膜(NCF)检验混合斑中的精斑 ABO 血型,但耗时。本文作者通过对此方法的改进,采用常彩琴等研制的抗人精特异蛋白血清(anti-human seminalpeculiar protein,ASPP),采用蛋清粘片热解离法检验混合斑中精斑 ABO 血型,耗时短,效果好。现介绍如下。     

自制与商业化抗人精液蛋白P30金标试剂条的法医学应用比较

目的　检测比较自制和商业销售的抗人精液蛋白P30 金标试剂条 ,探讨它们法医学应用的可靠性。方法　以人精液和人前列腺特异蛋白检测效价及灵敏度 ,以人阴道分泌物、人血清、人唾液和人初乳检测特异性 ,以保存不同年限的陈旧人精斑和斑痕类检材测定检验实际检材的能力。结果　自制抗人精液蛋白P30 金标试剂条效价及灵敏度与商业销售的试剂条中较好的基本相当 ,特异性则相当或好于商业试剂条 ,对非特异性测定均为阴性 ,而商业化试剂条则多有非特异性反应。结论　使用商业金标试剂条 ,要确定其是否合乎法医学检验标准与要求。     

gamma-Glutamyltransferase test as a new tool for identification of seminal stains

A simple qualitative method for identification of seminal stains based on a high activity of gamma-glutamyltransferase (gamma-GTP) in human semen is described. It employs the release of alpha-naphthylamine from N-gamma-glutamyl-alpha-naphthylamide by the gamma-GTP action: alpha-naphthylamine couples with Fast Garnet GBC salt to produce a strong brownish-red color. The data on its simplicity, specificity, and stability show that the present method is suitable for medicolegal examination of seminal stains as a preliminary test.     

Electronic data processing latex immunoassay for the identification of human seminal stains

Electronic data processing (EDP) latex immunoassay using anti-human seminal acid phosphatase (anti-HSAP) immune serum was applied for the species and organ identification of human seminal stains. This test proved to be highly sensitive and specific.     

用斑点ELISA方法检测p30确证人类精斑

作者建立了检测精浆特异性抗原p30的斑点ELISA试验方法,确证人类精斑。p30最小检出量为0.4ng。用该法盲测了室温存放1年的生物性斑痕180例,无一例假阳性和假阴性,证明用此法确证人类精斑的敏感性和特异性均优于传统的方法,具有操作简便、快速,检材用量小,结果准确等优点。     

人精浆蛋白双向谱型特征及其器官特异性的研究

本文应用双向聚丙烯酰胺凝胶电泳（2D—PAGE）技术分析了60例正常人精浆中的蛋白成分。阐明了人精浆蛋白双向电泳谱型的特征，并将人精浆蛋白双向电泳谱型与其他体液蛋白双向谱型进行了比较，证明人精浆蛋白双向电泳谱型具有器官特异性。本文还用2D－PAGE对不同条件下保存的精斑进行了认定。并将其应用于强奸案的鉴定。     

应用抗人精液单克隆抗体检验混合斑中精液成份的ABH血型物质

用本文报道的方法检测了123份混合斑检材（新鲜混合斑60份，陈旧混合斑63份），对保存在一年内的混合斑中精液成份中的血型物质检出率可达100％。     

Detection of human seminal γ-glutamyl transpeptidase in stains using sandwich ELISA

A sensitive and specific sandwich ELISA for human seminal γ-glutamyl transpeptidase (γ-GTP) was developed using a combination of monoclonal antibodies, SG1 and SG3, which we produced. For semen identification in forensic samples, we modified the assay so as to be more sensitive and to establish efficient extracting conditions. After testing the extracting abilities of several detergents, CHAPS and deoxy-BIGCHAP were chosen as the solubilizer. Polystyrene beads coated with SG1 were incubated with samples extracted by the detergents, and further with biotinylated SG3, followed by peroxidase-labeled streptavidin. γ-GTP was detected only in seminal samples. The sensitivity of this assay was 0.01 ng/ml of seminal γ-GTP equivalent to 10⁷ times diluted semen, which was ten times as compared with the previous plate assay. No significant seminal γ-GTP was detected in other biological stains such as blood, saliva and vaginal smear. The extract of a 500 fold diluted seminal stain, 8 months old, showed the detection limit. Seminal γ-GTP was detectable even in 14-year-old stains.     

一种改良酸性磷酸酶试验鉴定精斑的方法

本文报道了一种改良的酸性磷酸酶试验方法。用该法鉴定精斑，可克服非特异性反应，排除其它体液和植物汁液的影响。     

Detection of saliva traces using test strips

The test strip Rapignost-Amylase (Behring) for the rapid determination of alpha-amylase in the urine is also suitable for the determination of salivary amylase in stains stored up to 6 weeks at room temperature. The stains are extracted with physiological saline (extraction time 30 min), then the application zone of the strip is wetted with the extract. Positive amylase-reaction is recognised as a reddish-violet colouration of the reaction zone. Biological stains with low amylase concentrations (urine semen, vaginal secretion, mucus) react amylase negative. The method is uncomplicated and can be completed within 30 min. The test strips are easily available and stable during storage. Therefore the determination of saliva with test strips should be preferred to the clinical methods if the storage times of the stain are not longer than 4-6 weeks. It is a suitable procedure to determine salivary stains for use in forensic biology.     

The detection of inherited enzyme polymorphism in semen (author's transl)]

Our investigation of the occurrence of the enzymes phosphoglucomutase (PGM), glutamate-pyruvate-transaminase (GPT), adenylatkinase (AK), adenosine-desaminase (ADA), and 6-phophogluconate-dehydrogenase (6-PGD) produced the following results: The phosphoglucomutase type was demonstrated in the most sperm samples and seminal stains in accordance with the corresponding blood type. This enzyme is rather stable and could still be demonstrated well in 1-month old stains. The glutamate-pyruvate-transaminase can only seldom be determined in semen and seminal stains. We only found the GPT 1 type, which is known to have usually the strongest activity. The adenylatekinase was demonstrable in the most fresh ejaculates (not older than 24 h) and in about half the seminal stains (not older than 7 days)--The AK--2-band gets weak with increasing lay days, which may lead to incorrect determinations. The adenosine-desaminase could not be determined in sperm. On the contrary, 6-phosphogluconate-dehydrogenase could be demonstrated in fresh semen samples and also partly in seminal stains up to 7 days. The demonstration of the enzymes did not depend in any system on the secretor type.     

Effect of water immersion on seminal stains on cotton cloth

Reports on identification of seminal stains and spermatozoa on washed clothing are available. However, their detection on such clothing seems to depend on the washing material and the procedure adopted. It is reported here that prolonged immersion in water does not affect the detection of stains and spermatozoa.Results of experiments on water-immersed cotton clothing from 12 to 144 hours of water immersion are presented here. It is seen that intact human spermatozoa could be detected on such material even at 120 hours.