A valuable adjunct to FNA diagnosis of papillary thyroid carcinoma: in-house PCR assay for BRAF T1799A (V600E).

摘要: The diagnostic approach to thyroid nodules generally starts with FNA cytology. However, approximately one-fifth of cytologic evaluations yield indeterminate cytological findings but only 20% cases nodule cytology have a cancer diagnosis, emphasizing the need for an effective ancillary test based on material help prevent unnecessary surgery. Detection BRAFV600E mutation, genetic signature papillary carcinoma (PTC) in provides invaluable adjunct overcome limitations There are many ways detect V600E, such as direct DNA sequencing, allele-specific PCR and hybridization-based colorimetric methods. In this study, newer simple method is presented that removes requirements sequencing special equipment commercial kits. Two forward primers including mutant sequence specific (F2), one common reverse (R) primer were optimized generate 241 bp fragment (F1R), internal control, 141 (F2R) denoting presence V600E. Sensitivity studies revealed assay capable detecting V600E even 1 ng DNA. Direct data F1R proved specificity assay. For validation multiplex assay, archival slides used group lesions PTC, follicular carcinoma, adenoma, Hashimoto thyroiditis, benign nodules. PCR-based study practical, inexpensive one-step BRAF T1796A mutation samples. Diagn. Cytopathol. 2011;39:424–427. © 2010 Wiley-Liss, Inc.