Korean J Endocr Surg.  2010 Mar;10(1):12-18. 10.16956/kjes.2010.10.1.12.

Diagnosis of Papillary Thyroid Cancer via Detection of BRAF Mutation on Fine Needle Aspiration Cytology Slides

• Park WS
• Lee KE
• Song JY
• Chung YS
• Kim HY
• Koh SH
• Youn YK

• KMID: 2375727
• DOI: http://doi.org/10.16956/kjes.2010.10.1.12

Abstract

PURPOSE
The prevalence rate of the BRAF mutation in papillary thyroid cancer (PTC) is as high as about 52 to 83% in Korea. Preoperative detection of BRAF mutation on fine needle aspiration cytology (FNAC) slides may help the surgeon make better therapeutic decisions. The present study aims to assess the feasibility of the mutant allele specific amplification (MASA) and restriction fragment length polymorphism (RFLP) method with using conventional FNAC slides and we also wanted to evaluate the clinical role of preoperatively detecting BRAF mutation.
METHODS
We extracted the genomic DNA from 59 FNAC slides and performed direct sequencing (DS) for detecting BRAF mutation. We could use only 17 slides for the MASA method and 6 slides for the RFLP method due to the shortage of extracted DNA. Additionally, we retrospectively analyzed the cases for which a histological diagnosis could be made.
RESULTS
Genomic DNA was extracted from 23 out of the 59 FNAC slides. The BRAF mutation status could be assessed via DS in 33 out of the 59 FNAC slides. The concordance between the MASA method and DS and the RFLP method and DS was 36.3% and 66.7% respectively. The positive and negative predictive value of the 13 indeterminate nodules was 87.5% and 20%, respectively. We could not find any association between the BRAF mutations and the alleged risk factors of PTC.
CONCLUSION
We believe that the purity and the amount of the DNA template must be increased to detect BRAF mutation with using a FNAC slide. Preoperative detection of the BRAF mutation on a FNAC slide may refine the cytological diagnosis, but the application of assessing BRAF mutation as a prognostic marker is debatable.

Keyword

Papillary thyroid cancer; BRAF mutation; Fine needle aspiration cytology

MeSH Terms

Alleles
Biopsy, Fine-Needle*
Diagnosis*
DNA
Korea
Methods
Polymorphism, Restriction Fragment Length
Prevalence
Retrospective Studies
Risk Factors
Thyroid Gland*
Thyroid Neoplasms*
DNA

Figure

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  Fig. 2 BRAF exon15 PCR amplification. Which samples have enough and pure DNA to be sequenced might not be determined on electrophoresis gel.

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  Fig. 3 MASA products showing low sensitivity for BRAF mutation. Faint bands on No. 1, 3, 16 and 17 indicate the presence of BRAF mutation (Underlined numbers of samples have been proved for positive BRAF mutation by direct sequencing).

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  Fig. 4 RFLP products showing insecure results for BRAF mutation (Underlined numbers of samples have been proved for positive BRAF mutation by direct sequencing).

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