發展高敏感度之競爭性等位基因特異性PCR檢驗方法以輔助診斷甲狀腺乳突癌

Development of a Highly Sensitive Competitive Allele-Specific Real-Time PCR Assay to Assist in the Diagnosis of Papillary Thyroid Carcinoma

內分泌系統癌症中以甲狀腺癌占率最高，而其中有八成為甲狀腺乳突癌。而甲狀腺癌症中以BRAF V600E基因核酸位點突變最常發生。本篇研究主要目的為設立CAST-PCR基因檢測方法針對BRAF V600E進行檢驗。我們共收集200個臨床石蠟包埋組織進行BRAF V600E檢測，檢驗效能分析結果顯示此檢驗方法之臨床檢驗敏感度為81.9％，臨床檢驗特異性為100％，陽性預測值為100％，陰性預測值為83.3％。使用乳突癌細胞株及質體分析之檢驗結果確認，此檢驗方法之最低偵測突變敏感度為1％。同步比對細針抽取細胞學檢查（fine-needle aspiration cytology, FNAC）及石蠟包埋檢體確認兩種檢體之檢驗結果一致。經由以上結果得知，我們所設立的CAST-PCR的檢驗效能佳，利用FNAC檢體即可執行檢驗，得以輔助臨床醫師能精準診斷甲狀腺乳突癌，使臨床治療方針更即時。

Thyroid cancer is the most common malignancy neoplasm in the endocrine system. Papillary thyroid carcinoma (PTC) account for above 80% of thyroid cancers. BRAF V600E is the most commonly mutated gene in thyroid cancer. BRAF V600E mutations were assessed by competitive allele-specific PCR (CAST-PCR) in 200 patients with confirmed papillary thyroid carcinoma or benign findings. The clinical sensitivity was 81.9% and the clinical specificity was 100 %. Positive predictive value was 100% and the negative predictive value was 80.9%. Detection of BRAF V600E by CAST-PCR method in analytical sensitivity was 1%. The BRAF V600E mutation result of paired fine-needle aspiration cytology (FNAC) and FFPE were highly concordant with each other. In this study, we find an efficacy, reliability, and reproducibility CAST-PCR assay to assist in the diagnosis of papillary thyroid carcinoma.