Multiplex high resolution melt (HRM) messenger RNA profiling assays for body fluid identification

Traditional body fluid identification methods use a variety of technologically diverse techniques that do not permit the identification of all body fluids. Definitive identification of the biological material present can be crucial to a fuller understanding of the circumstances pertaining to a crime. Thus definitive molecular based strategies for the conclusive identification of forensically relevant biological fluids need to be developed. Messenger (mRNA) profiling is an example of such a molecular based approach.Current mRNA body fluid identification assays typically involve either capillary electrophoresis (CE) or quantitative RT-PCR (qRT-PCR) platforms, each with its own limitations. Both platforms require the use of expensive fluorescently labeled primers or probes. CE-based assays require separate amplification and detection steps thus increasing the time required for analysis. For qRT-PCR assays, only 3 or 4 markers can be included in a single reaction since each requires a different fluorescent dye. To simplify mRNA profiling assays and to reduce the time and cost of analysis, we have developed multiplex high resolution melt (HRM) assays that provide an identification of all forensically relevant biological fluids and tissues.     

蛇毒检测方法的实验研究

目的建立系统的蛇毒检测方法。方法通过对3种常见蛇毒样品检测,确定用ELISA法检测蛇毒的最适条件。结果本研究所建立的ELISA蛇毒检测法灵敏度为3.9ng/ml;交叉实验结果显示,该方法特异性高,只有个别蛇毒存在交叉反应;样本从采集到检出时间在1h40min之内;动物实验显示,半致死量中毒家兔的血液蛇毒检测在中毒后48h内有效,中毒死亡家兔的血液蛇毒检测在死后72h内有效。结论所建立方法灵敏、准确、快速、简便,是一种可靠的蛇毒检测法。     

How to regulate forensic familial DNA searching in Hungary?

Forensic DNA analysis has the potential to provide useful information for criminal justice even in cases where there is no match, neither between the DNA profile generated from the crime scene and the existing DNA profiles in criminal databases, nor between the DNA collected at a crime scene and potential suspects. In contrast to traditional forensic genetic testing, forensic familial DNA searching does not provide evidence, but helps to generate investigative leads and narrow down the range of potential offenders. The aim of this study is to examine, whether there is a need for special regulation of this topic in Hungary.     

Integrity verification of user space code

We present a novel approach for the construction and application of cryptographic hashes to user space memory for the purposes of verifying the provenance of code in memory images. Several key aspects of Windows behaviour which influence this process are examined in-depth. Our approach is implemented and evaluated on a selection of malware samples with user space components as well as a collection of common Windows applications. The results demonstrate that our approach is highly effective at reducing the amount of memory requiring manual analysis, highlighting the presence of malicious code in all the malware sampled.     

血液、脑脊液中尼可刹米的HPLC法分析

目的建立血液、脑脊液中尼可刹米的HPLC检测方法。方法取空白血液或脑脊液添加不同量的尼可刹米对照液,碱化后用二氯甲烷等有机溶剂提取,用HPLC方法检测。对方法的萃取回收率、线性、精密度等进行考察。结果方法选用二氯甲烷作为萃取溶剂,萃取回收率大于60%。血液中尼可刹米的在0.5~100μg/ml范围内线性良好,R2为0.9993;脑脊液中尼可刹米的线性范围为0.1~100μg/ml,R2为0.9996,最低检出限是5ng(S/N≥3)。检测血液和脑脊液中的尼可刹米方法的精密度均小于6%。结论所建方法准确、简便、灵敏度高,适用于法医毒物分析和临床药物分析。     

人血浆中尼莫地平气相色谱分析方法

建立人血液中尼莫地平的气相色谱分析方法 ,扩大药物检测范围及检测手段 ,以适应法医学鉴定的需要。以NaNO2 为氧化剂 ,将尼莫地平完全氧化成其吡啶衍生物 (PA)后用GC ECD进行分析。以标准尼莫地平对方法的线性范围、精密度进行了测试 ;以人血浆标准添加尼莫地平对样品处理方法、回收率进行考察。所建方法的线性范围是 1 2 1～ 2 4 2ng·ml 1(γ =0 9993) ;最低检出限为 1 0mg·ml 1(S/N =3) ;日内与日间的变异系数分别为 (5 77± 2 31) %与 (5 5 3± 0 70 ) % (n =4) ;平均回收率为 91 0 %～ 99 9%。该方法可用于尼莫地平血药浓度监测及法医学鉴定。     

测定单根毛发中吗啡含量的放射免疫方法

目的 建立测定单根毛发中吗啡含量的放免方法。方法 用卵清蛋白-琥珀酰吗啡作免疫原,免疫新西兰白兔获得高品质抗血清;HPLC纯化~(125)Ⅰ-吗啡,建立放射免疫方法,测定正常人和吸毒人员单根毛发。结果 抗体亲和常数为3.25×10~(11)L/M,放化纯度为95％,比放射性112μCi/μg;方法的灵敏度为0.01ng/ml。对5例正常人及5例戒毒所吸毒人员的单根毛发进行了检测。单根毛发长度9～24cm,重量为0.7～2.1mg,5例正常人测值为1.75±0.37ng/mg(x±s);5例吸毒人员测值为471±204ng/mg(x±s)。结论 所建方法可准确定量单根毛发中吗啡的含量。     

固相微萃取-气相色谱质谱法测定血浆中的氯氮平浓度

目的建立固相微萃取-气相色谱质谱法测定人血浆中氯氮平浓度的方法。方法以固相微萃取法提取血浆中的氯氮平,萃取头为100μm聚二甲基硅氧烷,洛沙平作内标,用气相色谱质谱选择离子法进行检测。结果本文建立的方法在5~2000 ng/ml浓度范围内呈线性关系,检测限为0.1 ng/ml(信噪比>3),低、中、高浓度(100、500、1000 ng/ml)平均相对回收率分别为98.6%、94.6%和94.6%,日内、日间RSD分别小于7.4%和7.1%。结论本文建立的固相微萃取-气相色谱质谱法灵敏度高、准确度好、操作简便,适用于氯氮平急性中毒案件的检测。     

颌面数字X线片性别判定指标的研究

目的筛选颌面侧位数字X线片(digital radiology,DR)的骨性定量指标,评估其在法医学性别判定中的应用价值。方法按纳入标准随机抽取颌面DR侧位片220例。对15项骨性指标(S-N,N-Me,Cd-Gn,S-Ns,S-Go(L),S-Gn,Cd-Go(L),SNA,SNB,SN-Npog,SN-MP,Gonial A,SN-RP,SN-NNs,SN-SGn)进行定量测量,采用方差分析方法分析各项指标是否有性别差异,应用逐步判别分析方法筛选出判别力较好的指标、建立判别函数并进行回代检验判别函数。结果200例颌面DR侧位片的15项指标中S-N,N-Me,Cd-Gn,S-Ns,S-Go(L),S-Gn,Cd-Go(L),SN-NNs等8项的性别差异有统计学意义(P<0.001);S-N,Cd-Gn,S-Ns,S-Gn 4项被纳入逐步判别函数。结论颌面DR侧位片中4项骨性指标(S-N,Cd-Gn,S-Ns,S-Gn)可用于性别判定,其判别准确率为95%。     

人线粒体DNA序列分析在法医学中的应用研究及其进展

综述人线粒体DNA(m tDNA)序列分析在法医学种属鉴别、个体识别,以及个体年龄推断中的应用研究及其进展,展望对m tDNA异质性的研究及建立人m tDNA数据库,并具有重要的法医学实践意义。     

IBD法在堂表亲缘关系鉴定中的应用

目的 使用血缘一致性(identity by descent,IBD)法计算堂表亲缘关系的堂表关系指数(first cousin index,FCI)和累积堂表关系指数(combined first cousin index,CFCI),为IBD法鉴定两个个体是否具有堂表亲缘关系提供科学手段.方法 取124对堂表兄弟姐...     

成人颅骨宽度的性别判别分析

为用颅骨判定性别提供依据。应用判别分析方法对收集东北地区已知性别成年男性和女性颅骨40具的颅骨宽度10个指标的性别差异进行分析。结果发现,10个指标中有7项性别差异显著(P<0.05),应用判别分析方法获得4个判别函数方程式,其判别率为96.3%～97.5%,本研究为可用颅骨判定性别提供新的依据。