鹌鹑减蛋综合征病毒单克隆抗体的制备及生物学特性

以鹌鹑减蛋综合征QAV C94病毒为抗原免疫BALB/c小鼠 ,制备单克隆抗体并对其生物学特性进行鉴定。获得 5株分泌单克隆抗体的细胞株 (命名为H1、H3、H4、H6和H8)。这 5株杂交瘤细胞可长期稳定地分泌单克隆抗体 ,分泌的单克隆抗体为IgG2a亚类 ,具有高度的特异性 ,只特异地作用于鹌鹑减蛋综合征病毒 ;微量血凝抑制试验显示H4有血凝抑制活性 ,腹水效价为 1∶2 6 ;这 5株单克隆抗体都没有沉淀活性 ,也不与CELO抗原发生沉淀反应 ;它们的亲和力以H4最大 ,相对亲和力依次为H4 >H6 >H1 >H8>H3。     

Proficiency testing (external quality assessment) of drug detection in oral fluid

Eighteen external quality assessment (proficiency testing) samples were prepared from client specimens collected with the Intercept^® oral fluid collection device and by spiking drug-free oral fluid. Samples were circulated in pairs at quarterly intervals to 13 UK and USA based laboratories for analysis by a panel of OraSure micro-plate Intercept^® enzyme immunoassay kits and hyphenated mass spectrophotometric techniques. During the survey, there was a single case of non-specificity in a false report for methadone. The major errors were of lack of sensitivity relative to the concentration thresholds specified for the immunoassays. The sensitivity for overall ‘present’/‘not found’ reports calculated as true positives/(true positives + false negatives) were for the amfetamine specific assay 50%, methyl-amfetamines 93%, barbiturates 64%, cannabinoids 73%, cocaine and metabolites 100%, benzodiazepines 69%, methadone 95%, opiates 79% (opiates excluding oxycodone 93%), phencyclidine 93% and human gamma-globulin 97%. A small number of the sensitivity errors were attributable to errors in chromatographic confirmation techniques.     

Determination of MDMA, MDA, MDEA and MBDB in oral fluid using high performance liquid chromatography with native fluorescence detection

This paper describes the analytical methodology for the determination of MDMA, MDA, MDEA and MBDB in oral fluid. After a liquid–liquid extraction, the analysis was carried out by high performance liquid chromatography (HPLC), with fluorescence detection. The detector wavelength was fixed at 285 nm for excitation and 320 nm for emission. The mobile phase, a mixture of phosphate buffer (pH = 5) and acetonitrile (75:25), and the column, Kromasil 100 C8 5 μm 250 mm × 4.6 mm, allowed good separation of the compounds in an isocratic mode in only 10 min. The method was validated and showed good limits of detection (2 ng/mL) and quantitation (10 ng/mL) for all the amphetamine derivatives. No interfering substances were detected. A stability study of these compounds in oral fluid stored at three different temperatures (−18, 4 and 20 °C) over 10 weeks was conducted, showing a time-dependent degradation of the four compounds.     

Analysis of Δ-tetrahydrocannabinol in oral fluid samples using solid-phase extraction and high-performance liquid chromatography–electrospray ionization mass spectrometry

An analytical method using solid-phase extraction (SPE) and high-performance liquid chromatography–mass spectrometry (LC–MS) has been developed and validated for the confirmation of Δ⁹-tetrahydrocannabinol (THC) in oral fluid samples. Oral fluid was extracted using Bond Elut LRC-Certify solid-phase extraction columns (10 cm³, 300 mg) and elution performed with n-hexane/ethyl acetate. Quantitation made use of the selected ion-recording mode (SIR) using the most abundant characteristic ion [THC + H⁺], m/z 315.31 and the fragment ion, m/z 193.13 for confirmation, and m/z 318.00 for the protonated internal standard, [d₃-THC + H⁺]. The method proved to be precise for THC, in terms of both intra-day and inter-day analyses, with coefficients of variation less than 10%, and the calculated extraction efficiencies for THC ranged from 76 to 83%. Calibration standards spiked with THC between 2 and 100 ng/mL showed a linear relationship (r² = 0.999). The method presented was applied to the oral fluid samples taken from the volunteers during the largest music event in Portugal, named Rock in Rio-Lisboa. Oral fluid was collected from 40 persons by expectoration and with Salivette^®. In 55% of the samples obtained by expectorating, THC was detected with concentration ranges from 1033 to 6552 ng/mL and in 45% of cases THC was detected at concentrations between 51 and 937 ng/mL. However, using Salivette^® collection, 26 of the 40 cases had an undetectable THC.     

法医物证学miRNA分析的研究进展

在种属和体液鉴定及降解检材等特殊案件的分析时,转录水平的miRNA所具有的生物属性及表达特点,使其能够发挥基因组DNA所不具备的价值。本文通过概述法医物证学miRNA研究的现状,对法医miRNA分析的研究策略和法医物证学应用前景进行了综述,以期为法医miRNA分析的应用研究提供借鉴。     

血色原微量分光法血痕确证试验

目的探讨血色原微量分光法作为血痕确证试验的可行性。方法用离心分离的检材浸洗液代替检材,与改良高山试剂进行血色原反应,用微量分光光度计测定吸收光谱,判断检材是否含血。在优化参数的基础上,用倍比稀释的粗制人血红蛋白测定方法的灵敏度,用常见疑似血液/血斑,以及不同保存时间、不同基质的血斑测定方法的特异性和检材适应能力。结果用血色原微量分光法,仅血液(斑)具有415nm、525nm和555nm三个吸收峰组成的特异光谱,与常见疑似血液/斑没有交叉。反应2min,上样2.5μL,稀释1000倍的人血可稳定获得三峰光谱。通过延长浸洗时间、设置基质对照,10年陈旧血纱、有色布上的血斑等均可有效确证。三个吸收峰的高度与检液的血含量呈正相关,且525nm和555nm的吸光度变化趋势一致(y=0.523 2x+0.027 4,R2=0.997 1)。结论血色原微量分光法灵敏度高、特异性强,快速简便,可纳入DNA检验流程之中,在实际检案中有较好的应用前景。替代传统结晶法用于教学,更符合当前学生技能和意识的培养要求。     

基于分子克隆方法制备标准分子量片段混合物

目的制备标准分子量DNA片段混合物。方法选取pMD18-T载体部分序列作为插入片段,设计引物,制备包含不同大小的标准分子量片段的克隆。大量培养细菌提取质粒,用双酶切、连接荧光接头的方法制备不同大小的带有荧光的标准分子量片段,混合,纯化,最终获得标准分子量片段混合物(内标)。结果用分子克隆方法制备出具有实用性的标准分子量片段混合物,其单个标准分子量片段大小分别为80bp、124bp、194bp、224bp、254bp、304bp、349bp、399bp、424bp、454bp。并且这些标准分子量片段混合物可以准确地对DNATyper15试剂盒的扩增产物进行检测。结论应用该方法制备了能满足研究和实验室要求的标准分子量片段混合物,为DNA分子量标准物标准品的制作提供了一种新的方法。     

人耳自动识别技术探讨

人耳识别是一种新的生物识别技术。一套完整的人耳自动识别系统一般应该包括以下几个环节:人耳图像采集、图像预处理、人耳图像的边缘检测与分割、特征提取、样本训练和模板匹配。人耳识别方法主要有主元分析法、神经网络法、力场转换方法、基于3D耳朵检测和识别方法。     

检测体液中氰含量的荧光光度法研究

本文报道了应用取代反应检测体液中氰含量的荧光光度法。探讨了检测的具体方法和有关条件对检测的影响。检测了正常人血、尿液和唾液中氰含量,并检测了小鼠氰化物染毒死亡后血中氰含量。研究表明,本法不但可检测体液中致死量的氰化物含量,亦可检测体液中正常氰含量。     

乡土秩序的失落——对流动人口犯罪的社会学思考

流动人口犯罪的原因及其防治机制不能从流动人口群体以外的范围内去寻找 ,必须深入到生成流动人口的具体社会结构中去探寻。该社会结构主要有二方面要素 :城乡二元格局 ;户籍管理体制和现存的庞大剩余劳动力。流动人口犯罪的基础性原因是存在于流动人口群体中的乡土秩序在城市中的失落 ,以及由此而产生的流动人口生存与发展预期的落空。要解决这些问题 ,必须寻找一个重建秩序之路 ,也就是经过小城镇化而达到全面的城市化。其目标是消除两种秩序的差别和对立 ,直至建立最大限度遏制流动人口犯罪的新型秩序。