Droplet digital PCR as an alternative to FISH for MYCN amplification detection in human neuroblastoma FFPE samples
Droplet digital PCR as an alternative to FISH for MYCN amplification detection in human neuroblastoma FFPE samples
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DOI：
10.1186/s12885-019-5306-0
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发表时间：
2019-01-28
期刊：
影响因子：
3.8
通讯作者：
Aravindan, Natarajan
中科院分区：
文献类型：
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作者：
Somasundaram, Dinesh Babu;Aravindan, Sheeja;Aravindan, Natarajan
背景资料：MYCN扩增与神经母细胞瘤的临床病程和患者生存率差直接相关，并作为最关键的阴性预后标志物。虽然荧光原位杂交（FISH）仍然是临床诊断神经母细胞瘤MYCN状态的金标准，但其局限性保证了快速、可靠、技术挑战性较小且廉价的替代方法的鉴定。方法：在本研究中，我们检查了液滴数字PCR的一致性，（ddPCR，结合免疫组织化学，IHC）与FISH用于在一组福尔马林固定石蜡包埋（FFPE）的人神经母细胞瘤样本中检测MYCN。在112例神经母细胞瘤病例中，ddPCR分析显示与FISH的一致性为96-100%。因此，IHC分级显示与FISH的一致性为92-100%。结论：ddPCR可用于NB患者MYCN扩增状态的检测，并可作为FFPE患者MYCN检测的一种技术性较低的方法。更重要的是，这些发现说明了FISH和ddPCR在MYCN状态检测中的一致性。总之，结果表明，快速，技术要求较低，廉价的ddPCR与IHC结合可以作为检测NB病例中MYCN状态的替代方法，其灵敏度与FISH几乎相同。
Background: MYCN amplification directly correlates with the clinical course of neuroblastoma and poor patient survival, and serves as the most critical negative prognostic marker. Although fluorescence in situ hybridization (FISH) remains the gold standard for clinical diagnosis of MYCN status in neuroblastoma, its limitations warrant the identification of rapid, reliable, less technically challenging, and inexpensive alternate approaches.Methods: In the present study, we examined the concordance of droplet digital PCR (ddPCR, in combination with immunohistochemistry, IHC) with FISH for MYCN detection in a panel of formalin-fixed paraffin-embedded (FFPE) human neuroblastoma samples.Results: In 112 neuroblastoma cases, ddPCR analysis demonstrated a 96-100% concordance with FISH. Consistently, IHC grading revealed 92-100% concordance with FISH. Comparing ddPCR with IHC, we observed a concordance of 95-98%.Conclusions: The results demonstrate that MYCN amplification status in NB cases can be assessed with ddPCR, and suggest that ddPCR could be a technically less challenging method of detecting MYCN status in FFPE specimens. More importantly, these findings illustrate the concordance between FISH and ddPCR in the detection of MYCN status. Together, the results suggest that rapid, less technically demanding, and inexpensive ddPCR in conjunction with IHC could serve as an alternate approach to detect MYCN status in NB cases, with near-identical sensitivity to that of FISH.