浅谈远程活体指纹信息登记查询工作站

远程活体指纹信息登记查询工作站是指纹自动识别网络系统的一个重要组成部分,连接着各级远程指纹工作站和违法犯罪人员的信息系统,是集信息采集、数据共享、远程传输和查询比对等多功能一体化的一种网络产品。使用该工作站可以大大提高基层公安机关采集指纹信息工作效率。     

Implementation of a robotized real-time PCR setup for the use of the Quantifiler™ Human DNA Quantification Kit

Human DNA quantification occupies a central role within the DNA analytic process of forensic casework samples as DNA quantification results have an important impact on the quality of the short tandem repeat data. Manual processing for the setup of quantification reactions can be time consuming and labor intensive. Therefore automation of quantitative real-time PCR setup was an important component of our DNA-analysis automation concept. Here we show the implementation of a robotized setup for the Quantifiler™ Human DNA Quantification Kit.     

批量陈旧血样DNA的自动化检验

目的建立批量陈旧血样DNA自动化提取检验的方法。方法采用普通磁珠法和本文建立的磁珠法经TECAN Freedom EVO150—8型自动化工作站分别提取540份陈旧血样模板DNA,采用Sinofiler^TM试剂盒进行荧光标记复合扩增。结果采用普通磁珠法和本文所建DNA提取方法,在540份样本中获得全部基因座STR分型的样本分别为217份和488份,检验成功率分别为40．2％和90．3％。结论本文所建方法可显著提高大批量陈旧血样自动化检验成功率。     

法医物证DNA自动化检验技术体系的研究

目的建立自动化工作站同步提取不同种类涉案法医生物检材DNA的新方法。方法选用TECAN Freedom EVO100．4、75—2型自动化提取、加样工作站,采用磁珠法及Chelex-100法对各类涉案生物检材进行DNA提取、PCR扩增、毛细管电泳检测其STR分型,进行比较测试。在“全国公安机关DNA数据库应用系统”中建立并应用实验室信息管理系统（LIMS）模拟实施规范化DNA检案。结果1552份各类检材,采用工作站-磁珠法提取DNA效果最佳,STR检测成功率为95％,工作站-Chelex法为88％;二者分别与其手工提取法比较,成功率无明显差异。92个样本同期检测,自动化工作站较手工操作DNA检案时间可缩减1．25倍。结论工作站域珠法提取涉案检材DNA,可获得满意的STR分型结果。应用LIMS管控,可有效防控污染,明显提高检案效率及鉴定质量。     

脱落上皮细胞类生物检材的自动化提取

目的建立一种自动化提取脱落上皮细胞类生物检材DNA的方法。方法附着于不同载体上的脱落上皮细胞类生物检材共278份,应用Eppendorf epMotion 5075LH工作站结合DNA IQTM系统提取模板DNA,并用Identifiler试剂盒进行STR检验。结果在278份被检的生物检材,其中126份检材获得13个基因座以上的STR分型,不同类型的检材其检出率不相同,最高达73.44%,最低为10.89%。结论脱落上皮细胞类检材应用自动化工作站提取DNA模板可在法医日常检案中广泛应用。     

胍盐裂解液对自动化提取工作站DNA回收率的影响

目的研究不同稀释度胍盐裂解液对自动化提取工作站DNA回收效率的影响。方法制备不同浓度的DNA样本,并将EQ1000法医DNA提取试剂盒中的胍盐裂解液梯度稀释,在本实验室配置的自动化提取工作站上,运行＂工作站磁珠法＂提取程序,AB-7500型荧光定量PCR仪检测提取前后的样本DNA浓度。选取2～7号梯度组DNA回收效率数据进行统计学检验,比较不同稀释度胍盐裂解液的处理对样品DNA回收效率的影响。结果 90%～60%稀释度胍盐裂解液组样品回收效率在66%左右;50%～20%稀释度组样品DNA回收效率在41%左右;稀释度在50%以下与60%以上组样品回收效率之间差异极显著。结论应用＂工作站磁珠法＂提取生物检材DNA,其DNA回收效率受胍盐裂解液浓度的影响。     

Automated DNA extraction using the QIAsymphony platform: Estimation of DNA recovery from simulated forensic stains

The aim of this study was to evaluate the forensic protocol recently developed by Qiagen for the QIAsymphony automated DNA extraction platform. Samples containing low amounts of DNA were specifically considered, since they represent the majority of samples processed in our laboratory. The analysis of simulated blood and saliva traces showed that the highest DNA yields were obtained with the maximal elution volume available for the forensic protocol, that is 200 μl. Resulting DNA extracts were too diluted for successful DNA profiling and required a concentration. This additional step is time consuming and potentially increases inversion and contamination risks. The 200 μl DNA extracts were concentrated to 25 μl, and the DNA recovery estimated with real-time PCR as well as with the percentage of SGM Plus alleles detected. Results using our manual protocol, based on the QIAamp DNA mini kit, and the automated protocol were comparable. Further tests will be conducted to determine more precisely DNA recovery, contamination risk and PCR inhibitors removal, once a definitive procedure, allowing the concentration of DNA extracts from low yield samples, will be available for the QIAsymphony.     

Adaptation and evaluation of the PrepFiler™ DNA extraction technology in an automated forensic DNA analysis process with emphasis on DNA yield, inhibitor removal and contamination security

Within the initial step of the forensic DNA analysis process, the DNA extraction efficiency and especially the removal of potential PCR inhibitors is crucial for subsequent steps, e.g. quantification by real-time PCR and amplification of short tandem repeats (STRs). The protocol of the PrepFiler™ Forensic DNA Extraction Kit was optimized for the application on a Tecan liquid handling workstation Freedom EVO^® 150. This modified application of the PrepFiler™ technology was compared with respect to DNA yield, sensitivity and the ability to remove potential PCR inhibitors to an established routine method working on the same liquid handling workstation based on ChargeSwitch^® Technology (CST) from Invitrogen.     

Successful validation of a fully automated sample lysis workstation

The Netherlands Forensic Institute recently validated a fully automated system for sample lysis, the AutoLys workstation and AutoLys tubes, developed in collaboration with Hamilton Robotics. We have carried out the developmental validation of the AutoLys system and concluded it offers a solution to drastically reduce user-errors, increase DNA yields, gain in productivity, while maintaining quality. This article reports on the study setup and discusses the results of comparing the AutoLys workstation to the conventional (manual) method. The AutoLys system proved to successfully process all commonly found forensic sample types, producing high quality extracts, containing on average 12.1% more DNA than manually extracted samples (as determined by qPCR and STR-profiling). The optimized robotic protocol minimizes the risk on cross-contamination (no cross-contamination detected in over 500 samples tested) and retains the flexibility to adjust chemically relevant variables specific to a laboratory's individual requirements. The fully automated process enables complete sample-tracking, drastically reducing the risk on user-errors. The NFI is currently validating an on-deck magnetic bead-based purification of the lysates.