Carturan Elisa, Tester David J, Brost Brian C, Basso Cristina, Thiene Gaetano, Ackerman Michael J
Department of Medico-diagnostic Sciences and Special Therapies, University of Padua Medical School, Padua, Italy.
Am J Clin Pathol. 2008 Mar;129(3):391-7. doi: 10.1309/VLA7TT9EQ05FFVN4.
One third of autopsy-negative sudden unexplained deaths (SUDs) can be attributed to a cardiac channelopathy. Typically, paraffin-embedded tissue (PET) is the only source of DNA available for genetic analyses. We examined different DNA extraction procedures, involving 2 deparaffinization methods, 2 digestion methods, 4 laboratory-based purification methods, and 5 commercial kits. Mutational analysis involving 25 RYR2 exons was performed on PET DNA from 35 SUD cases to evaluate the feasibility of using PET DNA for genetic testing. With the best PET-DNA extraction method, an average of only two thirds of the region of interest could be evaluated. Although we initially identified 5 missense mutations in 5 of 35 SUD cases, repeated analysis failed to confirm these mutations. DNA from PET should be considered error prone and unreliable in comprehensive surveillance of SUD-associated genes. Given these shortcomings, the standard autopsy for SUD should include archiving EDTA-preserved blood or frozen tissue to facilitate postmortem genetic testing.
三分之一的尸检阴性不明原因猝死（SUD）可归因于心脏离子通道病。通常，石蜡包埋组织（PET）是可用于基因分析的唯一DNA来源。我们研究了不同的DNA提取程序，包括2种脱石蜡方法、2种消化方法、4种基于实验室的纯化方法和5种商业试剂盒。对35例SUD病例的PET DNA进行了涉及25个RYR2外显子的突变分析，以评估使用PET DNA进行基因检测的可行性。采用最佳的PET-DNA提取方法，平均只能评估三分之二的感兴趣区域。虽然我们最初在35例SUD病例中的5例中鉴定出5个错义突变，但重复分析未能证实这些突变。在SUD相关基因的综合监测中，PET中的DNA应被视为容易出错且不可靠。鉴于这些缺点，SUD的标准尸检应包括存档EDTA保存的血液或冷冻组织，以方便死后基因检测。
