Riccardi Laura N, Lanzellotto Rossana, Falconi Mirella, Ceccardi Stefania, Bini Carla, Pelotti Susi
Department of Medical and Surgical Sciences, Institute of Legal Medicine, University of Bologna, via Irnerio, 49, 40126, Bologna, Italy.
J Forensic Sci. 2014 May;59(3):690-5. doi: 10.1111/1556-4029.12358. Epub 2013 Dec 6.
CYP2D6 polymorphism analysis is gaining increasing interest in forensic pharmacogenetics. Nevertheless, DNA recovered from forensic samples could be of poor quality and not suitable for long polymerase chain reaction required to type CYP2D6 gene prior to SNaPshot minisequencing analysis performed to define alleles with different enzymatic activity. We developed and validated following the guidelines of the Scientific Working Group on DNA Analysis Methods a tetraplex PCR yielding four amplicons of 597, 803, 1142, and 1659 bp encompassing the entire CYP2D6 gene to analyze eleven SNP positions by SNaPshot minisequencing. Concordance, sensitivity, and specificity were assessed. The method, applied to thirty-two forensic samples failed to amplify with long PCR, allowed the amplification of CYP2D6 gene in 62.5% of degraded samples. The new tetraplex PCR appears a suitable method for CYP2D6 analysis in forensic pharmacogenetics.
细胞色素P450 2D6（CYP2D6）基因多态性分析在法医药物遗传学领域正日益受到关注。然而，从法医样本中提取的DNA质量可能较差，不适用于在进行SNaPshot微测序分析以确定具有不同酶活性的等位基因之前对CYP2D6基因进行分型所需的长聚合酶链反应。我们按照DNA分析方法科学工作组的指南开发并验证了一种四重PCR，可产生包含整个CYP2D6基因的597、803、1142和1659 bp的四个扩增子，以便通过SNaPshot微测序分析11个单核苷酸多态性（SNP）位点。评估了一致性、灵敏度和特异性。该方法应用于32个无法通过长PCR扩增的法医样本，在62.5%的降解样本中成功扩增了CYP2D6基因。这种新的四重PCR似乎是法医药物遗传学中CYP2D6分析的一种合适方法。
