Evaluation of the Anyplex BRAF V600E Real-Time Detection Assay Using Dual-Priming Oligonucleotide Technology in Fine-Needle Aspirates of Thyroid Nodules
Annals of Laboratory Medicine
;
: 624-629, 2015.
Artigo
em Inglês
| WPRIM
| ID: wpr-76931
ABSTRACT
BACKGROUND:
Several molecular assays have been developed to detect the BRAF V600E mutation in fine needle aspirates (FNAs) for the diagnosis of papillary thyroid cancer. Using a multiplex PCR technique, we evaluated the Anyplex BRAF V600E Real-time Detection (Anyplex) assay and compared its efficacy with that of the Seeplex BRAF V600E ACE Detection (Seeplex) method.METHODS:
We tested 258 consecutive FNA specimens using the Seeplex and Anyplex assays. Any conflicting results between the two assays were confirmed by using mutant enrichment with 3'-modified oligonucleotide (MEMO) sequencing. The limits of detection (LODs) and reproducibility for each assay were evaluated with serially diluted DNA from a BRAF V600E-positive cell line.RESULTS:
The BRAF V600E mutation was detected in 36.4% (94/258) FNA specimens by either the Seeplex or Anyplex assay. Results for the two assays showed 93.4% (241/258) agreement, with a kappa value of 0.861 (95% confidence interval, 0.798-0.923). Of the eight specimens that were BRAF V600E-positive by the Anyplex assay but not by the Seeplex assay, five were found to be BRAF V600E-positive by MEMO sequencing. The mutation detection rate of the Seeplex and Anyplex assays was 79.0% and 84.0%, respectively, in the FNA specimens diagnosed as malignant (n=81). The LOD as determined by probit analysis was 0.046% (95% confidence interval, 0.019-0.532%).CONCLUSIONS:
The Anyplex assay performed better than the Seeplex assay with respect to the detection of the BRAF V600E mutation.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Oligonucleotídeos
/
DNA
/
Análise Mutacional de DNA
/
Nódulo da Glândula Tireoide
/
Primers do DNA
/
Polimorfismo de Nucleotídeo Único
/
Biópsia por Agulha Fina
/
Povo Asiático
/
Proteínas Proto-Oncogênicas B-raf
/
República da Coreia
Tipo de estudo:
Estudo diagnóstico
Limite:
Adulto
/
Idoso
/
Feminino
/
Humanos
/
Masculino
País/Região como assunto:
Ásia
Idioma:
Inglês
Revista:
Annals of Laboratory Medicine
Ano de publicação:
2015
Tipo de documento:
Artigo
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