Taqman Genotyping Assays Can Be Used on Decalcified and Paraffin-Embedded Tissue From Patients With Osteosarcoma
Taqman Genotyping Assays Can Be Used on Decalcified and Paraffin-Embedded Tissue From Patients With Osteosarcoma
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Taqman 基因分型测定可用于骨肉瘤患者的脱钙组织和石蜡包埋组织
DOI：
10.1002/pbc.22654
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发表时间：
2011-01-01
影响因子：
3.2
通讯作者：
Hoogerbrugge, Peter M.
中科院分区：
文献类型：
--
作者：
Hagleitner, Melanie M.;Coenen, Marieke J. H.;Hoogerbrugge, Peter M.
背景对于骨肉瘤等5年总生存率为50- 60%的癌症，存档的组织学标本可以成为生物材料的有用来源，但这种材料通常已经脱钙并用福尔马林固定多年。在我们的研究中，我们研究了从这些组织中获得的DNA是否可以用于可靠的单核苷酸多态性（SNP），基因分型方法我们使用Taqman SNP基因分型分析研究了药物转运蛋白MDR 1中的两个SNP。将来自20名骨肉瘤存活患者新鲜分离DNA的生殖系DNA的基因型与来自脱钙福尔马林固定的存档材料的基因型进行比较，相同患者的石蜡包埋（FFPE）块结果与从外周血细胞提取的DNA相比，脱钙的FFPE衍生的DNA产生更小的PCR片段，具有200 bp的可靠大小。然而，在本研究的20个病例中，我们能够评估19个病例的每个SNP。所有成功的基因分型样本均显示100%从FFPE组织DNA中获得的基因型与从同一患者血液DNA中获得的基因型之间的一致性结论总之，我们已经证明脱钙FFPE组织可以用于使用Taqman（R）的遗传多态性分析。等位基因鉴别分析这形成了一个独特的机会，将联合收割机在遗传研究中的新见解与历史患者队列相结合儿科血液癌症2011 56 35-38（C）2010 Wiley利斯公司
Background In cancers like osteosarcoma with a 5 year over all survival of 50-60%, archived histological specimens can be a useful source of biological material However this material generally has been decalcified and formalin fixed for many years In our study, we investigated whether DNA obtained from these tissues can be used for reliable single nucleotide polymorphism (SNP) genotyping Procedure We studied two SNPs in the drug transporter MDR1 using Taqman (R) SNP genotyping assays Genotypes of the germ line DNA derived from freshly isolated DNA of 20 surviving patients with osteosarcoma were compared with genotypes obtained from archived material from decalcified formalin fixed, paraffin embedded (FFPE) blocks of the same patients Results Decalcified FFPE derived DNA yielded smaller PCR fragments compared to DNA extracted from peripheral blood cells, with a reliable size of 200 bp However, we were able to evaluate each SNP in 19 of 20 cases included in this study All successfully genotyped samples showed 100% concordance between genotypes obtained from DNA of FFPE tissue and the genotypes obtained from DNA of blood from the same patients Conclusions In conclusion we have demonstrated that decalcified FFPE tissue can be used for genetic polymorphism analysis using Taqman (R) allelic discrimination assays This forms a unique opportunity to combine new insights in genetic research with historical patient cohorts Pediatr Blood Cancer 2011 56 35-38 (C) 2010 Wiley Liss Inc