Sanchez J J, Børsting C, Balogh K, Berger B, Bogus M, Butler J M, Carracedo A, Court D Syndercombe, Dixon L A, Filipović B, Fondevila M, Gill P, Harrison C D, Hohoff C, Huel R, Ludes B, Parson W, Parsons T J, Petkovski E, Phillips C, Schmitter H, Schneider P M, Vallone P M, Morling N
Section of Forensic Genetics, Department of Forensic Medicine, Faculty of Health Sciences, University of Copenhagen, 11 Frederik V's Vej, DK-2100 Copenhagen, Denmark.
Forensic Sci Int Genet. 2008 Jun;2(3):176-83. doi: 10.1016/j.fsigen.2007.12.002. Epub 2008 Feb 11.
We report the results of an inter-laboratory exercise on typing of autosomal single nucleotide polymorphisms (SNP) for forensic genetic investigations in crime cases. The European DNA Profiling Group (EDNAP), a working group under the International Society for Forensic Genetics (ISFG), organised the exercise. A total of 11 European and one US forensic genetic laboratories tested a subset of a 52 SNP-multiplex PCR kit developed by the SNPforID consortium. The 52 SNP-multiplex kit amplifies 52 DNA fragments with 52 autosomal SNP loci in one multiplex PCR. The 52 SNPs are detected in two separate single base extension (SBE) multiplex reactions with 29 and 23 SNPs, respectively, using SNaPshot kit, capillary electrophoresis and multicolour fluorescence detection. For practical reasons, only the 29 SBE multiplex reaction was carried out by the participating laboratories. A total of 11 bloodstains on FTA cards including a sample of poor quality and a negative control were sent to the laboratories together with the essential reagents for the initial multiplex PCR and the multiplex SBE reaction. The total SNP locus dropout rate was 2.8% and more than 50% of the dropouts were observed with the poor quality sample. The overall rate of discrepant SNP allele assignments was 2.0%. Two laboratories reported 60% of all the discrepancies. Two laboratories reported all 29 SNP alleles in all 10 positive samples correctly. The results of the collaborative exercise were surprisingly good and demonstrate that SNP typing with SBE, capillary electrophoresis and multicolour detection methods can be developed for forensic genetics.
我们报告了一项针对犯罪案件法医遗传学调查中常染色体单核苷酸多态性（SNP）分型的实验室间比对结果。欧洲DNA分析小组（EDNAP），国际法医遗传学协会（ISFG）下属的一个工作组，组织了此次比对。共有11个欧洲和1个美国法医遗传学实验室对由SNPforID联盟开发的一个包含52个SNP的多重PCR试剂盒中的一个子集进行了检测。这个包含52个SNP的多重试剂盒在一次多重PCR中扩增52个具有常染色体SNP位点的DNA片段。使用SNaPshot试剂盒、毛细管电泳和多色荧光检测，在两个单独的单碱基延伸（SBE）多重反应中分别检测这52个SNP，其中一个反应检测29个SNP，另一个检测23个SNP。出于实际原因，参与实验室仅进行了包含29个SNP的SBE多重反应。总共11份FTA卡上的血迹样本，包括一份质量较差的样本和一个阴性对照，与用于初始多重PCR和多重SBE反应的必需试剂一起被送至各实验室。总的SNP位点丢失率为2.8%，且超过50%的丢失情况出现在质量较差的样本中。SNP等位基因分型不一致的总体发生率为2.0%。两个实验室报告了所有差异的60%。两个实验室在所有10个阳性样本中正确报告了所有29个SNP等位基因。此次协作比对的结果出奇地好，表明使用SBE、毛细管电泳和多色检测方法进行SNP分型可应用于法医遗传学。
