TY - JOUR
T1 - Droplet digital PCR as an alternative to FISH for MYCN amplification detection in human neuroblastoma FFPE samples
AU - Somasundaram, Dinesh Babu
AU - Aravindan, Sheeja
AU - Yu, Zhongxin
AU - Jayaraman, Muralidharan
AU - Tran, Ngoc T.B.
AU - Li, Shibo
AU - Herman, Terence S.
AU - Aravindan, Natarajan
N1 - Funding Information:
The authors are supported by research funding from the National Institutes of Health (NIH 1P20GM103639–01) Centers of Biomedical Research Excellence (COBRE) Program and the University of Oklahoma Health Sciences Center (OUHSC) Department of Radiation Oncology Research Development Funds.
Funding Information:
The authors acknowledge the neuroblastoma specimen providers: Department of Pathology, University of Oklahoma Health Sciences Center; Cooperative Human Tissue Network (CHTN), which is funded by the National Cancer Institute (NCI), and; Oregon Health and Science University Biospecimen core. The authors acknowledge the Stephenson Cancer Center (SCC) – COBRE Histochemistry and immunology core, SCC-COBRE Biospeci-men pathology core, and OUHSC Pediatrics Clinical Genetics Core - Cytogenetic Molecular division for their services. The authors also acknowledge the OUHSC Staff Editor (Ms. Kathy Kyler) for the help in critically reviewing this manuscript.
Publisher Copyright:
© 2019 The Author(s).
PY - 2019/1/28
Y1 - 2019/1/28
N2 - 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.
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.
KW - ddPCR
KW - FISH
KW - Immunohistochemistry
KW - MYCN amplification
KW - Neuroblastoma
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U2 - 10.1186/s12885-019-5306-0
DO - 10.1186/s12885-019-5306-0
M3 - Article
C2 - 30691436
AN - SCOPUS:85060650520
VL - 19
JO - BMC Cancer
JF - BMC Cancer
SN - 1471-2407
IS - 1
M1 - 106
ER -