交叉免疫电泳对血清痕进行个人识别的实验研究

血痕的个人识别是法医学实践中最重要的课题之一。目前虽然有多种血型系统包括红细胞血型、红细胞酶型,血清型、以及白细胞血型等应用于血痕的个人识别,但每个系统都只能进行排除,而不能认定。交叉免疫电泳又名双向定量免疫电泳,由于其具有不需要复杂设备,操作简单、所需样品量少、特异性较强及重复性好等许多     

用酶标抗人Hb单克隆抗体检测微量血痕种属的实验

法医学血痕种属检验的方法很多，如环状沉淀试验，琼脂糖单扩散试验以及对流免疫电泳试验等。这些方法操作简便，但灵敏度低，易受血痕所附着的客体以及化学合成物等因素的干扰。笔者参照文献［1］介绍的方法，用酶标抗人Hb单克隆抗体检测微量血痕种属，取得良好的效果，现报道如下。材料与方法一、材料酶标扰人Hb单克隆抗体和8管反应板（华能公司博塞试剂研究所生产）；抗人Hb单克隆抗体（白求恩国际和平医院提供）；抗人Hb多克隆抗体（公安部第二研究所血清室提供）；三羟甲基氨基甲烷等试剂（北京化学试剂厂）；牛血清白蛋白（BSA）（…     

免疫固定法同步检测C3表型及其转化率

本文利用醋酸纤维素薄膜电泳免疫固定法结合光密度扫描，同步检测C3表型及其转化率，与交叉免疫电泳比较，本方法对不同区带的分辨力更高，定量更精确，并且快速、简便、灵敏。     

抗人精液血清特异性的研究

确证精斑常用抗人精液血清作沉淀反应。这种抗血清特异性差,常与人类某些体液发生交叉反应,以致精斑确证试验结果不可靠,影响鉴定结果的准确性。作者用人类精子、精浆、精液与人类精浆特异性抗原p30分别免疫家兔,制备各种抗血清,用环状沉淀、琼脂双向扩散及免疫电泳等技术对这些抗血清的特异性进行了研究,报道如下。     

人精浆特异蛋白P_30的分离纯化与鉴定

本文介绍人精浆特异蛋白P_(30)分离纯化及鉴定的方法。人精浆经蒸馏水透析后经Sephadex G100、G150和G75三种不同的柱层析即可获得精浆特异蛋白。SDS-PAGE、免疫电泳、免疫双扩散证明其纯度和特异性均符合要求。它与已知抗P_(30)血清呈强阳性反应,用其免疫制备的抗血清和已知抗P_(30)血清,在免疫电泳中只与精浆或精浆特异蛋白形成一条完全相同的沉淀线,SDS-PAGE测得分子量约为30,000。故笔者将其亦定名为P_(30)。     

Gc蛋白的分离纯化

本文报道Gc蛋白的分离与纯化.人血浆经DEAE-Sephadex-A50, Sephadex G100和DEAE-Sephadex-A50三次柱层析即可获得纯化Gc.纯化Gc经PAGE, SDS-PAGE和免疫电泳等证明,其纯度和特异性均符合要求.     

运用不连续性对流免疫电泳试验进行血痕的种属鉴定

对流免疫电泳技术在法医学上,用于血痕种属鉴别,已得广泛运用,但仅限于连续电泳,即槽缓冲液的类型和离子强度,与琼脂板的相同。本文作者运用了不连续电泳,即槽缓冲液的类型和离子强度,与琼脂板的不一样,进行了血痕种属鉴别试验,认为本法具有形成的沉淀线清晰,灵敏度高等优点,很有实用价值。     

Gc亚型在鉴定亲子关系中的应用

1959年Hirschfeld用免疫电泳方法研究人血清α_2—球蛋白时,发现一些与Hp无关的沉淀带,根据其迁移率可以分为快速、中速和慢速三种。这类蛋白被称为型特异性成分Gc(Group-Specific Compon-ent),受控于常染色体上的两个共显性等位基因Gc~1和Gc~2。而后进一步研究发现Gc的主要生理功能是结合并传递维生素D,故又被称为维生素D结合蛋白。     

抗GC血清的制备及鉴定

作者用分离纯化的GC蛋白免疫新西兰兔制备了抗GC血清。鉴定结果表明,抗GC血清与进口商品抗GC血清特异性相同;与人血清其他蛋白没有交叉反应。琼脂双向扩散法测得抗血清效价为128;可检出GC蛋白的最低浓度为3.1μg/ml。免疫电泳法可测出入血清三种GC表型。     

用免疫电泳技术检测人血痕中Gc的表型

<正> 在60年代初,有人尝试用免疫电泳技术对人血痕进行 Gc 定型,均以血痕中的Gc 蛋白在电泳时,向阳极飘移而告失败。Shinomiya 和 Baelen 发现 Gc 蛋白飘移的原因是与其血痕中破碎细胞释放出来的肌动蛋白结合形成复合物所致。用4M 盐酸胍处理血痕,可使复合物解聚,恢复 Gc 的正常电泳迁移率。     

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。     

型特异性沉淀素血清检验人唾液斑精斑ABO血型的研究与应用

本文介绍了利用型特异性沉淀素血清环状沉淀法检验人唾液斑、精液斑ABO血型的方法与实验结果,并与中和试验及解离试验进行了比较。实验结果表明,本法操作简便,对多种干扰条件下的唾液斑、精斑均具有高度的型特异性,并能从分泌液与血液的混合斑中准确地鉴别出分泌液的血型。本法仅需0.4cm的分泌斑纱线即可进行血型鉴定,其灵敏度高于中和试验而略低于热解离试验,并能有效地检出陈旧分泌液斑中的型物质,因此适于在实际检案中应用。     

ELISA检测精浆特异蛋白P_(30)鉴定人精斑

<正> 1978年美国Sensabaugh分离出精浆特异蛋白P30,并成功的制备出相应的抗P30血清。1987年我国也研制出抗人精浆特异蛋白P30血清,并应用该种抗血清建立了几种琼脂扩散和电泳方法鉴定人精斑。由于人精液中P30含量不高,平均为1.52±0.676mg/ml,鉴定微量精斑存在困难。为了更有效的提高方法的灵敏度。我们建立了鉴定微量人精斑的ELISA固相P30抗体法,现报告如下。     

An evaluation of gamma-glutamyl transpeptidase (GGT) and p30 determinations for the identification of semen on postcoital vaginal swabs

The following study entails the investigation of gamma-glutamyl transpeptidase (GGT) and p30 for the identification of seminal stains in sexual assault cases. A commercial kit was used to test for GGT activity, while p30 was demonstrated with a crossed electrophoresis technique. Specificity, sensitivity, and stability of both markers were studied. Postcoital swabs from lab staff were tested for GGT and p30. In addition, 144 postcoital swabs from case material were tested for p30, spermatozoa, and acid phosphatase. Results show p30 to be a useful semen marker particularly in cases of azoospermia. However, GGT was found to be unsuitable for forensic science casework.     

HLA—A基因座分步PCR—SSP分型研究

目的使HLA基因分型能应用于法医常见检材的个人识别。方法 建立检测HLA—A基因座的分步PCR—SSP方法。先用一对HLA—A基因座特异的引物作第一次扩增,以所得产物为模板,分别用对HLA—A30、A31、A33特异的3对引物作第二次扩增,二次扩增的产物经电泳判型。结果 1130例血清分型为HLA—A30、A31、A33的血痕,其PCR—SSP分型和血清分型的不符合率为29％;室温保存2年的精斑、唾液斑,保存18年的血痕第一次扩增均获得满意的结果。结论法医亲子鉴定和个人识别宜用基因分型替代血清分型。HLA—A基因座分步PCR—SSP基因分型适用于法医检材。     

抗人N单克隆抗体的理化特性研究及其初步应用

本文报告理化因素对抗人N单克隆抗体(ON_1D_3)活性的影响。P~H及反应温度对ON_1D_3单克隆抗体与人红细胞的血凝反应有显著的影响。较适宜的实验条件为P~H7.0,温度30℃。在此条件下对155例新鲜血液及60例陈旧血痕样品进行MN分型。结果与用市售兔抗N血清测定的结果完全一致,证明在上述条件下,ON_1D_3单克隆抗体可用于临床常规血液分型及法医学上对陈旧血痕MN血型的测定。     

用胶乳凝集抑制试验鉴定人类精斑

作者用人类精浆致敏的胶乳和人初乳吸收后的抗人精液血清,采用胶乳凝集抑制试验凹玻板法盲测了180份生物性斑痕,结果表明此法确证人类精斑的敏感性,准确性均高于精子检出法,而与抗 p30血清琼脂双向扩散结果一致,且可用于混合斑的检验,具有快速、准确、简便等优点。     

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.     

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 acid phosphatase test two minute cut-off: An insufficient time to detect some semen stains

The ability to detect semen in sexual offence cases is a crucial first step to locating stains which may be suitable for DNA profiling. Since the development of the acid phosphatase test in the late1950s by Stuart S. Kind, the process undertaken to perform the test has gone largely unchanged. The method currently accepted by operational forensic science laboratories allows 2 min for a reaction to be obtained, and until relatively recently, this has not been challenged. In this research, samples of semen were obtained from three donors and a range of dilutions for each sample were prepared. Each dilution was subjected to acid phosphatase testing using both direct testing and the ‘press test’ method. The results showed that semen could be detected in excess of 15 min in dilutions up to 1 in 400 using the press test method and in dilutions up to 1 in 1000 using the direct method. Of further significance was the observation that using the press test method, the two minute cut-off was insufficient to detect the majority of stains and in some cases, semen stains as strong as 1 in 20 dilutions. This research provides compelling evidence for protocols currently utilised in forensic practice to be reviewed in order that forensic scientists do not overlook potential evidential material that may prove suitable for body fluid identification such as DNA STR profiling.