Droplet digital polymerase chain reaction assay and peptide nucleic acid‐locked nucleic acid clamp method for RHOA mutation detection in angioimmunoblastic T‐cell lymphoma
Autor: | Hirotake Wakamatsu, Mitsunobu Shimadzu, Kosei Matsue, Yasuhito Suehara, Mamiko Sakata-Yanagimoto, Kota Fukumoto, Keiichiro Hattori, Daisuke Komori, Manabu Fujisawa, Sharna Tanzima Nuhat, Manabu Kusakabe, Shigeru Chiba |
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Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
0301 basic medicine
Peptide Nucleic Acids Cancer Research Angioimmunoblastic T-cell lymphoma RHOA next‐generation sequencing (NGS) Oligonucleotides PNA‐LNA clamp method Polymerase Chain Reaction law.invention 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine law medicine Humans Digital polymerase chain reaction Locked nucleic acid Genetics Genomics and Proteomics droplet digital polymerase chain reaction (ddPCR) assay Polymerase chain reaction Peptide nucleic acid biology Oligonucleotide High-Throughput Nucleotide Sequencing Lymphoma T-Cell Peripheral General Medicine Original Articles medicine.disease Molecular biology 030104 developmental biology Oncology chemistry G17V RHOA mutation 030220 oncology & carcinogenesis Immunoblastic Lymphadenopathy Mutation Nucleic acid biology.protein Original Article rhoA GTP-Binding Protein angioimmunoblastic T‐cell lymphoma |
Zdroj: | Cancer Science |
ISSN: | 1349-7006 1347-9032 |
Popis: | Angioimmunoblastic T‐cell lymphoma (AITL) is a subtype of nodal peripheral T‐cell lymphoma (PTCL). Somatic RHOA mutations, most frequently found at the hotspot site c.50G > T, p.Gly17Val (G17V RHOA mutation) are a genetic hallmark of AITL. Detection of the G17V RHOA mutations assists prompt and appropriate diagnosis of AITL. However, an optimal detection method for the G17V RHOA mutation remains to be elucidated. We compared the sensitivity and concordance of next‐generation sequencing (NGS), droplet digital PCR (ddPCR) and peptide nucleic acid‐locked nucleic acid (PNA‐LNA) clamp method for detecting the G17V RHOA mutation. G17V RHOA mutations were identified in 27 of 67 (40.3%) PTCL samples using NGS. ddPCR and PNA‐LNA clamp method both detected G17V mutations in 4 samples in addition to those detected with NGS (31 of 67, 46.3%). Additionally, variant allele frequencies with ddPCR and those with NGS showed high concordance (P T;50G > T], p.Gly17Leu in PTCL198; c.[50G > T;51A > C], p.Gly17Val in PTCL216; and c.50G > A, p.Gly17Glu in PTCL223) were detected using NGS. These sequence changes could not appropriately be detected using the ddPCR assay and the PNA‐LNA clamp method although both indicated that the samples might have mutations. In total, 34 out of 67 PTCL samples (50.7%) had RHOA mutations at the p.Gly17 position. In conclusion, our results suggested that a combination of ddPCR/PNA‐LNA clamp methods and NGS are best method to assist the diagnosis of AITL by detecting RHOA mutations at the p.Gly17 position. |
Databáze: | OpenAIRE |
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