PMID- 30691436 OWN - NLM STAT- MEDLINE DCOM- 20190503 LR - 20200225 IS - 1471-2407 (Electronic) IS - 1471-2407 (Linking) VI - 19 IP - 1 DP - 2019 Jan 28 TI - Droplet digital PCR as an alternative to FISH for MYCN amplification detection in human neuroblastoma FFPE samples. PG - 106 LID - 10.1186/s12885-019-5306-0 [doi] LID - 106 AB - 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. FAU - Somasundaram, Dinesh Babu AU - Somasundaram DB AD - Departments of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Boulevard, Oklahoma City, OK, 73104, USA. FAU - Aravindan, Sheeja AU - Aravindan S AD - Stephenson Cancer Center, Oklahoma City, OK, USA. FAU - Yu, Zhongxin AU - Yu Z AD - Department of Pathology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Boulevard, BMSB 451, Oklahoma City, OK, 73104, USA. FAU - Jayaraman, Muralidharan AU - Jayaraman M AD - Stephenson Cancer Center, Oklahoma City, OK, USA. AD - Department of Cell Biology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Boulevard, BMSB 553, Oklahoma City, OK, 73104, USA. FAU - Tran, Ngoc T B AU - Tran NTB AD - Department of Pathology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Boulevard, BMSB 451, Oklahoma City, OK, 73104, USA. FAU - Li, Shibo AU - Li S AD - Department of Pediatrics, University of Oklahoma Health Sciences Center, 1200 Children's Ave. Ste 14000, Oklahoma City, OK, 73104, USA. FAU - Herman, Terence S AU - Herman TS AD - Departments of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Boulevard, Oklahoma City, OK, 73104, USA. AD - Stephenson Cancer Center, Oklahoma City, OK, USA. FAU - Aravindan, Natarajan AU - Aravindan N AUID- ORCID: 0000-0001-9150-3911 AD - Departments of Radiation Oncology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Boulevard, Oklahoma City, OK, 73104, USA. naravind@ouhsc.edu. AD - Department of Pathology, University of Oklahoma Health Sciences Center, 940 Stanton L. Young Boulevard, BMSB 451, Oklahoma City, OK, 73104, USA. naravind@ouhsc.edu. AD - Department of Anesthesiology, University of Oklahoma Health Sciences Center, 920 SL Young Blvd #1140, Oklahoma City, OK, 73104-5036, USA. naravind@ouhsc.edu. LA - eng GR - P20 GM103639/GM/NIGMS NIH HHS/United States GR - P30 CA225520/CA/NCI NIH HHS/United States GR - NIH 1P20GM103639-01/NH/NIH HHS/United States PT - Comparative Study PT - Journal Article DEP - 20190128 PL - England TA - BMC Cancer JT - BMC cancer JID - 100967800 RN - 0 (Biomarkers, Tumor) RN - 0 (MYCN protein, human) RN - 0 (N-Myc Proto-Oncogene Protein) RN - 1HG84L3525 (Formaldehyde) SB - IM MH - Biomarkers, Tumor/genetics MH - Formaldehyde MH - Gene Amplification MH - Humans MH - Immunohistochemistry MH - In Situ Hybridization, Fluorescence MH - N-Myc Proto-Oncogene Protein/*genetics MH - Neuroblastoma/*diagnosis/genetics/pathology MH - Paraffin Embedding MH - *Polymerase Chain Reaction MH - Sensitivity and Specificity PMC - PMC6348625 OTO - NOTNLM OT - FISH OT - Immunohistochemistry OT - MYCN amplification OT - Neuroblastoma OT - ddPCR COIS- ETHICS APPROVAL AND CONSENT TO PARTICIPATE: All protocols were approved by the University of Oklahoma Health Sciences Center Institutional Review Board with permission for the research use of de-identified specimens (OUHSC IRB #6207). All experiments were performed in accordance with the University of Oklahoma Health Sciences Institutional Review Board guidelines and regulations for the protection of human subjects. CONSENT FOR PUBLICATION: Not Applicable. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2019/01/30 06:00 MHDA- 2019/05/06 06:00 PMCR- 2019/01/28 CRDT- 2019/01/30 06:00 PHST- 2017/12/13 00:00 [received] PHST- 2019/01/16 00:00 [accepted] PHST- 2019/01/30 06:00 [entrez] PHST- 2019/01/30 06:00 [pubmed] PHST- 2019/05/06 06:00 [medline] PHST- 2019/01/28 00:00 [pmc-release] AID - 10.1186/s12885-019-5306-0 [pii] AID - 5306 [pii] AID - 10.1186/s12885-019-5306-0 [doi] PST - epublish SO - BMC Cancer. 2019 Jan 28;19(1):106. doi: 10.1186/s12885-019-5306-0.