RNA/DNA co-analysis from blood stains-Results of a second collaborative EDNAP exercise
RNA/DNA co-analysis from blood stains-Results of a second collaborative EDNAP exercise
复制标题
DOI：
10.1016/j.fsigen.2011.02.004
复制
发表时间：
2012-01-01
影响因子：
3.1
通讯作者：
Ballantyne, J.
中科院分区：
文献类型：
--
作者：
Haas, C.;Hanson, E.;Ballantyne, J.
第二次关于RNA/DNA共同分析用于体液鉴定和STR图谱分析的协作活动是由欧洲DNA图谱分析小组组织的。参与实验室使用RNA/DNA共提取或单独RNA提取方法分析了6份人血斑、2份血液稀释系列（5-0.001 μ l血液）和（可选）人或非人来源的真实或模拟病例样本。两种新的mRNA复合物用于鉴定血液：高度敏感的双链体（HBA，HBB）和中等敏感的五链体（ALAS 2，CD 3G，ANK 1，SPTB和PBGD）。实验室使用不同的化学品和仪器。所有18个参与实验室都能够成功分离和检测干血迹中的mRNA。13个实验室同时提取RNA和DNA从个人染色，并能够利用mRNA分析，以确认血液的存在，并获得常染色体STR的血迹捐赠者。从旧的和受损的个案样本中也获得了血液和高质量DNA图谱的阳性鉴定。使用不同的分析策略，该方法被证明是重现性和灵敏度。这种涉及RNA/DNA共提取策略的协作练习的结果支持基于mRNA的系统用于与当前DNA分析方法兼容的法医案例中的血液鉴定的潜在用途。(C)2011爱思唯尔爱尔兰有限公司保留所有权利。
A second collaborative exercise on RNA/DNA co-analysis for body fluid identification and STR profiling was organized by the European DNA Profiling Group (EDNAP). Six human blood stains, two blood dilution series (5-0.001 mu l blood) and, optionally, bona fide or mock casework samples of human or non-human origin were analyzed by the participating laboratories using a RNA/DNA co-extraction or solely RNA extraction method. Two novel mRNA multiplexes were used for the identification of blood: a highly sensitive duplex (HBA, HBB) and a moderately sensitive pentaplex (ALAS2, CD3G, ANK1, SPTB and PBGD). The laboratories used different chemistries and instrumentation. All of the 18 participating laboratories were able to successfully isolate and detect mRNA in dried blood stains. Thirteen laboratories simultaneously extracted RNA and DNA from individual stains and were able to utilize mRNA profiling to confirm the presence of blood and to obtain autosomal STR profiles from the blood stain donors. The positive identification of blood and good quality DNA profiles were also obtained from old and compromised casework samples. The method proved to be reproducible and sensitive using different analysis strategies. The results of this collaborative exercise involving a RNA/DNA co-extraction strategy support the potential use of an mRNA based system for the identification of blood in forensic casework that is compatible with current DNA analysis methodology. (C) 2011 Elsevier Ireland Ltd. All rights reserved.