Bongiovanni M, Molinari F, Eszlinger M, Paschke R, Barizzi J, Merlo E, Giovanella L, Fasolini F, Cattaneo F, Ramelli F, Mazzucchelli L, Frattini M
Institute of Pathology, Locarno, Switzerland.
Division of Endocrinology and Nephrology, University of Leipzig, Leipzig, Germany.
Cytopathology. 2015 Oct;26(5):288-96. doi: 10.1111/cyt.12226. Epub 2014 Dec 8.
The application of molecular tests to thyroid fine needle aspiration (FNA) has been shown to be a valuable tool to better refine the pre-operative malignant risk of patients with indeterminate cytology results. In this study, we investigated the feasibility of using the laser capture microdissection (LCM) technique to obtain DNA and RNA for molecular tests in routine thyroid FNA smears.
Nine coupled FNA and histological retrospective cases and 31 prospective FNA cases with a follicular neoplasm/suspicious for a follicular neoplasm (FN/SFN) diagnosis were included in this study. Both cytological and histological specimens were investigated by direct sequencing and reverse transcription-polymerase chain reaction (RT-PCR) for BRAF and RAS mutations and for PAX8/PPARG and RET/PTC rearrangements, respectively.
LCM yielded good DNA and RNA quality in all cases (100%) in both series, irrespective of the staining used (Giemsa, Papanicolaou, immunostain for thyroglobulin) and the cytology technique (conventional or liquid-based preparations). Total mutations found in the FNA and in the corresponding histological specimen in both series were: one PAX8/PPARG rearrangement in a follicular carcinoma (FC), four NRAS mutations [in two FCs, one papillary carcinoma and one follicular adenoma (FA)] and one HRAS mutation in one FA. The sensitivity was 67% and the specificity was 91%.
LCM is a valuable tool to obtain good quality DNA and RNA for molecular tests in cytological material from thyroid FNA, and can be a useful option in the management of patients with an FN/SFN FNA diagnosis.
分子检测应用于甲状腺细针穿刺活检(FNA)已被证明是一种有价值的工具,可更好地细化细胞学结果不确定患者的术前恶性风险。在本研究中,我们调查了使用激光捕获显微切割(LCM)技术从常规甲状腺FNA涂片获取用于分子检测的DNA和RNA的可行性。
本研究纳入了9例FNA与组织学配对的回顾性病例以及31例诊断为滤泡性肿瘤/可疑滤泡性肿瘤(FN/SFN)的前瞻性FNA病例。分别通过直接测序和逆转录聚合酶链反应(RT-PCR)对细胞学和组织学标本进行BRAF和RAS突变以及PAX8/PPARG和RET/PTC重排的检测。
在两个系列的所有病例(100%)中,无论使用何种染色方法(吉姆萨染色、巴氏染色、甲状腺球蛋白免疫染色)和细胞学技术(传统涂片或液基制片),LCM均能获得高质量的DNA和RNA。两个系列中FNA及其相应组织学标本中发现的总突变情况如下:1例滤泡癌(FC)中有1个PAX8/PPARG重排,4例NRAS突变[2例FC、1例乳头状癌和1例滤泡性腺瘤(FA)],1例FA中有1个HRAS突变。敏感性为67%,特异性为91%。
LCM是从甲状腺FNA的细胞学材料中获取用于分子检测的高质量DNA和RNA的有价值工具,对于FN/SFN FNA诊断的患者管理可能是一个有用的选择。