骨髓活检标本基因突变检测方法的比较。
Comparison of methodologies for the detection of mutations in bone marrow trephine specimens.
机构信息
Medical School, University of Oxford, Oxford, UK.
Nuffield Division of Clinical Laboratory Sciences, University of Oxford, Oxford, UK
出版信息
J Clin Pathol. 2019 Jun;72(6):406-411. doi: 10.1136/jclinpath-2019-205734. Epub 2019 Mar 14.
AIMS
V600E detection assists in the diagnosis of hairy cell leukaemia (HCL); however, testing practices vary. We evaluated the clinical utility of 5 mutation testing strategies for use on bone marrow trephines (BMT).
METHODS
11 HCL, 5 HCL 'mimic', 2 treated HCL and 10 normal BMT specimens were tested for mutant comparing Sanger sequencing, pyrosequencing, amplicon-based next generation sequencing (NGS), automated (Idylla) PCR and immunohistochemistry (IHC).
RESULTS
PCR and IHC were cheaper and identified V600E in 100 % of HCL cases. Pyrosequencing detected the mutation in 91%, NGS in 55% of cases and Sanger sequencing in 27%. All assays gave wild-type results in HCL mimics and normal BMT samples.
CONCLUSIONS
PCR and IHC were most sensitive and cost-effective, but these have limited scope for multiplexing and are likely to be replaced by NGS gene panels or whole genome sequencing in the medium to long term.
目的
V600E 检测有助于诊断毛细胞白血病（HCL）；然而，检测实践存在差异。我们评估了 5 种突变检测策略在骨髓活检（BMT）中的临床应用价值。
方法
对 11 例 HCL、5 例 HCL“模拟”病例、2 例治疗后的 HCL 病例和 10 例正常 BMT 标本进行了突变检测，比较了 Sanger 测序、焦磷酸测序、基于扩增子的下一代测序（NGS）、自动化（Idylla）PCR 和免疫组织化学（IHC）。
结果
PCR 和 IHC 更便宜，可识别 100%的 HCL 病例中的 V600E。焦磷酸测序检测到 91%的病例存在突变，NGS 检测到 55%的病例存在突变，Sanger 测序检测到 27%的病例存在突变。所有检测方法在 HCL 模拟病例和正常 BMT 样本中均未检测到野生型 。
结论
PCR 和 IHC 最敏感、最具成本效益，但这些方法在多重检测方面的应用范围有限，可能会被 NGS 基因面板或全基因组测序所取代，在中短期内取代它们。
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