Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas.
ALK is an established causative oncogenic driver in neuroblastoma, and is likely to emerge as a routine biomarker in neuroblastoma diagnostics. At present, the optimal strategy for clinical diagnostic evaluation of ALK protein, genomic and hotspot mutation status is not well-studied. We evaluated AL...
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doaj-657f55be47104141b121d2c64e824ed32020-11-24T21:42:52ZengPublic Library of Science (PLoS)PLoS ONE1932-62032014-01-0199e10657510.1371/journal.pone.0106575Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas.Benedict YanChik Hong KuickMalcolm LimKavita VenkataramanChandana TennakoonEva LohDerrick LianMay Ying LeongManikandan LakshmananVinay TergaonkarWing-Kin SungShui Yen SohKenneth T E ChangALK is an established causative oncogenic driver in neuroblastoma, and is likely to emerge as a routine biomarker in neuroblastoma diagnostics. At present, the optimal strategy for clinical diagnostic evaluation of ALK protein, genomic and hotspot mutation status is not well-studied. We evaluated ALK immunohistochemical (IHC) protein expression using three different antibodies (ALK1, 5A4 and D5F3 clones), ALK genomic status using single-color chromogenic in situ hybridization (CISH), and ALK hotspot mutation status using conventional Sanger sequencing and a next-generation sequencing platform (Ion Torrent Personal Genome Machine (IT-PGM)), in archival formalin-fixed, paraffin-embedded neuroblastoma samples. We found a significant difference in IHC results using the three different antibodies, with the highest percentage of positive cases seen on D5F3 immunohistochemistry. Correlation with ALK genomic and hotspot mutational status revealed that the majority of D5F3 ALK-positive cases did not possess either ALK genomic amplification or hotspot mutations. Comparison of sequencing platforms showed a perfect correlation between conventional Sanger and IT-PGM sequencing. Our findings suggest that D5F3 immunohistochemistry, single-color CISH and IT-PGM sequencing are suitable assays for evaluation of ALK status in future neuroblastoma clinical trials.http://europepmc.org/articles/PMC4154751?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Benedict Yan Chik Hong Kuick Malcolm Lim Kavita Venkataraman Chandana Tennakoon Eva Loh Derrick Lian May Ying Leong Manikandan Lakshmanan Vinay Tergaonkar Wing-Kin Sung Shui Yen Soh Kenneth T E Chang |
spellingShingle |
Benedict Yan Chik Hong Kuick Malcolm Lim Kavita Venkataraman Chandana Tennakoon Eva Loh Derrick Lian May Ying Leong Manikandan Lakshmanan Vinay Tergaonkar Wing-Kin Sung Shui Yen Soh Kenneth T E Chang Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas. PLoS ONE |
author_facet |
Benedict Yan Chik Hong Kuick Malcolm Lim Kavita Venkataraman Chandana Tennakoon Eva Loh Derrick Lian May Ying Leong Manikandan Lakshmanan Vinay Tergaonkar Wing-Kin Sung Shui Yen Soh Kenneth T E Chang |
author_sort |
Benedict Yan |
title |
Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas. |
title_short |
Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas. |
title_full |
Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas. |
title_fullStr |
Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas. |
title_full_unstemmed |
Platform comparison for evaluation of ALK protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas. |
title_sort |
platform comparison for evaluation of alk protein immunohistochemical expression, genomic copy number and hotspot mutation status in neuroblastomas. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2014-01-01 |
description |
ALK is an established causative oncogenic driver in neuroblastoma, and is likely to emerge as a routine biomarker in neuroblastoma diagnostics. At present, the optimal strategy for clinical diagnostic evaluation of ALK protein, genomic and hotspot mutation status is not well-studied. We evaluated ALK immunohistochemical (IHC) protein expression using three different antibodies (ALK1, 5A4 and D5F3 clones), ALK genomic status using single-color chromogenic in situ hybridization (CISH), and ALK hotspot mutation status using conventional Sanger sequencing and a next-generation sequencing platform (Ion Torrent Personal Genome Machine (IT-PGM)), in archival formalin-fixed, paraffin-embedded neuroblastoma samples. We found a significant difference in IHC results using the three different antibodies, with the highest percentage of positive cases seen on D5F3 immunohistochemistry. Correlation with ALK genomic and hotspot mutational status revealed that the majority of D5F3 ALK-positive cases did not possess either ALK genomic amplification or hotspot mutations. Comparison of sequencing platforms showed a perfect correlation between conventional Sanger and IT-PGM sequencing. Our findings suggest that D5F3 immunohistochemistry, single-color CISH and IT-PGM sequencing are suitable assays for evaluation of ALK status in future neuroblastoma clinical trials. |
url |
http://europepmc.org/articles/PMC4154751?pdf=render |
work_keys_str_mv |
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