Medical School, University of Oxford, Oxford, UK.
Nuffield Division of Clinical Laboratory Sciences, University of Oxford, Oxford, UK
J Clin Pathol. 2019 Jun;72(6):406-411. doi: 10.1136/jclinpath-2019-205734. Epub 2019 Mar 14.
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).
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).
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.
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.
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。
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）。
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 。
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。
