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.
Article
in En
| WPRIM
| ID: wpr-76931
Responsible library:
WPRO
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.
Key words
Full text:
1
Index:
WPRIM
Main subject:
Oligonucleotides
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DNA
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DNA Mutational Analysis
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Thyroid Nodule
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DNA Primers
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Polymorphism, Single Nucleotide
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Biopsy, Fine-Needle
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Asian People
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Proto-Oncogene Proteins B-raf
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Republic of Korea
Type of study:
Diagnostic_studies
Limits:
Adult
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Aged
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Female
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Humans
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Male
Country/Region as subject:
Asia
Language:
En
Journal:
Annals of Laboratory Medicine
Year:
2015
Type:
Article