G3M T typing by dot immunobinding with monoclonal anti-G3M T antibody

Dilutions of 100 serum samples of various GM phenotypes were dotted onto a nitrocellulose membrane. The serum dot-blots were detected with peroxidase-labeled anti-G3M T monoclonal antibody (anti-G3M T MAb). Up to a 1 : 256 dilution could be G3M T-typed correctly. By use of anti-G3M T MAb and peroxidase-labeled anti-mouse IgG or the biotin-avidin system instead of use of labeled anti-G3M T MAb, up to a 1 : 512 or 1 : 1024, respectively, dilution was typable. As with previous work with G3M G MAb, the dot immunobinding (DIB) method for G3M T typing was found very simple and practical.     

Failure to detect antihuman-immunoglobulinallotype-active antibodies in diagnostic sera directed against some enterobacteria

Diagnostic sera to determine antigenic properties of bacteria were tested to clarify the question whether these sera also contain antibodies being active against human immunoglobulin allotypes. Sera directed against various strains of Escherichia coli, Salmonella, and Shigella were found to be negative for anti-Ig-allotype activity.     

抗人同种异型G1m(3)单克隆抗体研制及特异性鉴定

用快速液相色谱仪从G1m（3）阳性人血浆中纯化IgG1蛋白，用其免疫BALB／C／小鼠．建立了一株分泌抗人G1m（3）单克隆抗体的细胞株（D7E8）。经抑制ELISA，直接ELISA及斑点ELISA分析，证明D7E8抗体具有G1m（3）单一特异性。其培养上清液效价为512倍，并初步应用于法医办案。     

胶体金免疫层析一步法快速检测G1m(3)因子

建立一种简便快速的胶体金免疫层析一步法 ,用于检测 G1m(3)因子。采用柠檬酸三钠还原法制备胶体金颗粒 ,标记抗人 G1m(3)单克隆抗体 ,研制出抗人 G1m(3)因子免疫层析检测试剂盒。样品的 G1m(3)因子 ,与测试条上的金标记抗体结合后沿着反应膜移动 ,再与膜上固相抗体结合形成肉眼可见的红色反应带。使用该方法可检出 10万倍稀释的血清样品 ,整个试验只需 5 min完成。对常见的 2 3种动物血 (痕 )检验 ,未出现交叉反应。在 10 0例样品的检测中 ,本方法的检测结果 ,与 Dot- EL ISA的检测结果的符合率为 10 0 %。该方法适用于法医物证快速检验。     

G3m(21) typing by ELISA and dot immunobinding with enzyme-labeled monoclonal anti-G3m(21) antibody

Simple, rapid methods are described for G3m(21) typing with peroxidase-labeled monoclonal anti-G3m(21) antibody. In G3m(21) typing by ELISA, microtiter wells were coated directly with the test antigen, which was detected with the enzyme-labeled monoclonal antibody. To further simplify the procedure, a dot immunobinding method was developed. The antigen in the test serum applied onto a nitrocellulose membrane was successfully detected with the enzyme-labeled monoclonal antibody. These methods, particularly the dot immunobinding, are suitable for forensic casework because they are rapid and simple and require no technical skill.     

Some aspects in preparation and use of wide spectrum antiglobulin sera in forensic medical studies

Wide spectrum (polyspecific) antiglobulin sera were obtained by rabbit immunization with 10% solution of zymosan-charged globulin (product of yeast digestion with trypsin) with complete Freund's adjuvant. The resultant sera meet the international standards and can be used for forensic medical identification of serum G1m(1) group and for detecting incomplete antierythrocytic complement-fixing anti-Daffi, anti-Kell, and other antibodies in blood transfusion service.     

抗丁丙诺啡单克隆抗体的制备

目的建立抗丁丙诺啡单克隆抗体的杂交瘤细胞株，制备高特异性的丁丙诺啡单克隆抗体，并对其免疫学特性进行鉴定。方法在丁丙诺啡的分子上连接活性羧基基团，通过缩合反应将丁丙诺啡半抗原连接于血蓝蛋白（KLH）和小牛血清白蛋白（BSA），形成完全抗原。以完全抗原免疫Balb／c小鼠，通过细胞融合，筛选等杂交瘤技术，建立稳定的分泌抗丁丙诺啡单克隆抗体的杂交瘤细胞株。通过腹腔注射杂交瘤细胞，诱导小鼠产生含有单抗的腹水。用辛酸-硫酸铵加亲和层析法纯化抗丁丙诺啡单克隆抗体。采用酶联免疫反应和胶体金膜层析实验测定丁丙诺啡单抗的特异性以及免疫反应动力学参数。结果共获得3株分泌抗丁丙诺啡单克隆抗体的杂交瘤细胞株，分别命名为7E6，6G4和3C2。7E6，6G4抗体灵敏度为10．0ng／ml，3C2抗体灵敏度为20．0ng／ml。7E6，6CA和3C2抗体的亲和常数分别为3．6×10^-9 mol／L，4．3×10^-9 mol／L和6．3×10^-9 mol／L。特异性测试结果表明7E6和6G4抗体与40种药物、毒品无任何交叉反应，而3C2抗体与吗啡有交叉反应。结论杂交瘤细胞株7E6和6G4产生的抗丁丙诺啡单克隆抗体具有很高的特异性和灵敏度。     

抗人转铁蛋白单链抗体的研制及部分性能研究

研制人转铁蛋白的单链抗体。以纯化人转铁蛋白免疫小鼠,制备小鼠脾脏 mRNA,RT-PCR 扩增重轻链可变区基因;Linker 连接及引入 Sfi Ⅰ和 NotⅠ酶切位点构建单链抗体基因;同载体 pCANTAB 5E 连接,转化 E.coliTG1细胞,以 M13K07挽救制备噬菌体抗体呈现文库,筛选抗原阳性重组噬菌体粒子(Panning);感染 E.coliHB2151菌株,制备可溶抗体并检测其性质;测定抗体基因序列并分析同源性及结构特征。制备出4株分泌抗人转铁蛋白可溶抗体的 E.coli HB2151菌株。抗体基因只由小鼠抗体的轻重链可变区基因构成,开放读码,无终止子,并紧接一末尾为一琥珀终止密码的 E 末端序列,其对应氨基酸序列 V_H 和 V_L 部分均遵从抗体可变区特征。实验证明,可把基因工程抗体技术应用于法医血清学,以制备新型抗体试剂。     

Determination of the blood group properties of bones subjected to the decay process

The aim of the present study was to make generalizations concerning the results of serological examinations of bones subjected to the process of decay. The specimens examined were taken from the femur. Some of the specimens had been subjected to decay for a period of some years, after which they were examined using the absorption-elution method in accordance with classical principles. Finally, successive dilutions of eluted antibodies were performed with a view to establishing the "antigenic force" of the material under examination. It was found that, owing to the process of bone decay, there is a decrease in the original "antigenic force", which is accompanied by the appearance (in a weak form) of non-specific serological reactions, which are much weaker in strength than specific ones. It was concluded that for the interpretation of the actual blood group of decayed bones the term "diagnostic" can only be applied to a range of dilutions not exceeding that of non-specific agglutinations.     

Gm typing by enzyme-linked immunosorbent assay (ELISA)

A solid-phase ELISA for Gm typing is described. A mixture of anti-Gm serum (or monoclonal anti-Gm antibody) and test serum was incubated in microtiter wells coated with IgG or its fragments of appropriate Gm type. After washing of the wells, the bound antibody was detected with peroxidase-labeled second antibody. The Glm(3), G3m(16), and G3m(21) antigens could be identified by this technique. Since some of the human anti-Gm sera and anti-Rh0 sera required for the conventional hemagglutination-inhibition method are hard to obtain, the ELISA system using anti-Gm antibodies and no anti-Rh0 sera may serve as an alternative to the conventional method.     

鼠抗人转铁蛋白单链抗体基因的表达

目的 制备抗人转铁蛋白单链抗体。方法 拼装好并带有酶切位点粘性末端的鼠抗人转铁蛋白的单链抗体基因经凝胶电泳定量后,同载体pCANTAB 5E连接,以其转化E.coli TG1细胞,经辅助噬菌体M13K07挽救构建噬菌体抗体表面呈现文库、抗原包被固相材料富集选择阳性重组噬菌体克隆（Panning）。感染能产生可溶抗体片段的大肠杆菌菌株E.coli HB2151,制备可溶抗体,以Anti-E末端抗体进行Western blot、ELISA检验和分子量测定。结果 人转铁蛋白的单链抗体基因成功表达。结论 用基因工程抗体技术制备抗体是可行的。     

Development and application of immunoassay for paraquat: radioimmunoassay

A sensitive radioimmunoassay for paraquat is reported. Anti-paraquat antisera were produced by repeated immunization in rabbits with 1-methyl, 1'-hexanoic acid-4,4'-bipyridinium (MHBP) coupled to bovine serum albumin (BSA). Less than 0.5 ng of paraquat dichloride was detectable by this assay system. These antisera were strongly cross-reactive with the bipyridyl ring and methyl group in either the 1- or 1'-position of paraquat. The determination of paraquat in tissues of paraquat-poisoned cadavers was also carried out.     

A dot-blot-immunoassay for semen identification using a polyclonal antibody against semenogelin, a powerful seminal marker

Among various seminal plasma proteins, semenogelin (Sg), produced in the seminal vesicle, has been considered a candidate for demonstrating the presence of semen. Sg consists of two proteins, one 52 kDa (Sg-I) in size, and the other a mixture of 71 and 76 kDa proteins (Sg-II). Recombinant Sg-I and Sg-II proteins were obtained using a baculovirus system and then injected into a rabbit to produce the respective antibodies [Characterization of recombinant precursor proteins of the human seminal plasma sperm motility inhibitor synthesized in insect cells, Int. J. Mol. Med. 2 (1998) 693]. When liquefied seminal plasma was immunoblotted with the anti-Sg-I and Sg-II antibodies, the anti-Sg-II antibody identified a wider range of the polypeptides originating from Sg than did the anti-Sg-I antibody. A dot-blot-immunoassay using anti-Sg-II antibody revealed a clear immunoreactive spot even when the semen was diluted 6400-fold. However, this assay showed that the Sg antigen was undetectable in saliva, urine, vaginal secretions, sweat, nasal secretions and serum. To determine the stability of Sg antigenic activity, filter paper with dried semen stains were kept at 37, 4 and 22 degrees C for 1, 6 and 18 months, respectively, and the Sg antigenic activity was examined. The activity was detectable in an area not less than 0.5 cm x 0.5 cm under all of the above environmental conditions during each period. Finally, semen was mixed with saliva or blood at various volumetric ratios, and used as a source of dried stains. The Sg antigenic activity was detectable in the stains until the ratio of semen to saliva or blood reached 1:8. These results suggest that Sg may be useful as a marker for semen identification.     

酶标2G8单克隆抗体斑点ELISA快速检验人血痕G2m(23)因子

利用自制的抗人 G2m(23)酶标2G8酶标单克隆抗体,首次应用直接斑点 ELISA 方法检测血痕中 G2m(23)因子。该法最小检出量为0.000048μl 血清;整个试验可在15分钟内完成。对常见的13种动物血和8种鱼血无交叉反应;盲测450例干血痕结果全部正确。该方法具有快速、准确、灵敏、简便等特点,有较大的实用价值。     

Preferential Selection Independent of Race and Gender: Effects on Self-Evaluations and Newcomer Information-Seeking Behaviors

The current study investigated the effects of an experimentally imposed program of preferential selection on beneficiary self-evaluations and newcomer information-seeking behavior. One hundred-twenty undergraduates were randomly assigned to a classification condition (in which they were informed that they tended to think in either an analytical or abstract manner) and collaborated on a task in groups of three. A fourth participant was introduced into each of these 40 extant groups under either a condition of preferential selection or not. Preferentially selected newcomers were shown to have more positive self-evaluations than their nonpreferentially selected counterparts. The presence or absence of a similar (in terms of thinking style) incumbent moderated the effect of being preferentially selected on the use of specific information-seeking behaviors.     

A screening method for urinary methamphetamine--latex agglutination inhibition reaction test

Methamphetamine in urine samples from abusers was detected by the latex agglutination inhibition reaction test with latex-antibody (Latex-Ab) and latex-methamphetamine (Latex-MA) reagents. Anti-methamphetamine antibody was produced in rabbits by immunization with bovine serum albumin (BSA)-methamphetamine conjugate. Latex particles were coated with antibodies or with rabbit serum albumin (RSA)-methamphetamine conjugate to obtain Latex-Ab and Latex-MA reagents, respectively. The results are read at 4-5 min after mixing the latex reagents. The sensitivity of this method for methamphetamine was 0.4 micrograms/ml urine. Methamphetamine analogs (methylephedrine, amphetamine, phentermine, methoxyphenamine, ephedrine, beta-phenylethylamine, OH-methamphetamine, OH-amphetamine, and OH-ephedrine) all cross-react in varying degrees, while glucosiurea and albuminurea give false positive results in the tests. Though attention must be paid to these effects this simple and rapid test is suitable for the mass screening of urine samples.     

Laboratory investigation of deaths due to anaphylaxis.

To establish a useful laboratory protocol to investigate possible cases of fatal anaphylaxis, we measured mast-cell-derived tryptase levels and allergen-specific immunoglobulin E (IgE) antibody levels in sera obtained prior to or within 24 h after death from 19 anaphylaxis victims. Elevated serum tryptase levels (range = 12 ng/mL to 150 micrograms/mL) were found in nine of nine Hymenoptera sting fatalities, six of eight food-induced fatalities, and two of two reactions to diagnostic therapeutic agents. Tryptase levels were normal (less than 10 ng/mL) in 57 sequential sera obtained postmortem from six control patients. Tryptase could not be measured in pleural or pericardial fluids for technical reasons. Serum IgE antibodies were elevated in five of the nine Hymenoptera sting fatalities and in eight of the eight fatal food reactions; assays were unavailable for the two diagnostic/therapeutic agents. If elevated, the victim's serum IgE antibodies to food could be used to identify allergens in uneaten portions of foods consumed shortly before the anaphylactic event. IgE antibodies were moderately stable during storage in a variety of anticoagulants at room temperature for up to 11 weeks. Elevated mast-cell-derived tryptase levels in postmortem sera reflect antemortem mast cell activation and may be used as a marker for fatal anaphylaxis. If assays are available for IgE antibodies to relevant allergens, such assays provide evidence for antemortem sensitization; these assays may be modified to identify allergens in foods consumed by victims of food-induced anaphylaxis.     

The Role of Site Variance in the American Judicature Society Field Study Comparing Simultaneous and Sequential Lineups

Objectives

Police departments often use photo lineups for eyewitness identification purposes. A widely adopted lineup reform designed to reduce eyewitness misidentifications involves switching from the standard simultaneous photo presentation format to a sequential format. These two lineup procedures were recently tested in the American Judicature Society (AJS) field study, which was conducted in four different police jurisdictions. The results from two phases of that investigation reached opposite conclusions as to which lineup procedure is superior, and the purpose of our current investigation was to elucidate the role of site variance in shaping those contrasting conclusions.

Methods

In previous analyses, the field study data were either (1) aggregated across all four study sites or (2) drawn from only one study site (Austin, Texas). Here, we analyze the data separately for the Austin study site, where 69 % of the eyewitnesses were tested, and the other three study sites combined, where 31 % of the eyewitnesses were tested.

Results

The results indicate significant site variance between the Austin and non-Austin study sites. In addition, the results suggest that aggregating the data across sites played a determinative role in creating the apparent disagreement about which lineup procedure is diagnostically superior.

Conclusions

Once large differences across the AJS study sites are taken into consideration, there is no longer any disagreement about which lineup procedure is superior. The simultaneous procedure is diagnostically superior to the sequential procedure, but the sequential procedure sometimes induces more conservative responding (a result that can and often does masquerade as diagnostic superiority).

     

The use of monoclonal anti-M and anti-N antibodies in forensic medicine

Anti-M and anti-N monoclonal antibodies (MA) may be useful for bloodstain analysis by absorption-elution reaction. In order to detect N antigen in bloodstains aged up to 4 weeks the material tested must be treated by methanol. The material fixation is not recommended for analysis of "aged" bloodstains as well as for M antigen detection. Anti-M MA may be used for analysis of liquid blood using agglutination reaction.