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
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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