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甲状腺结节细针穿刺活检作为分子研究的可能来源:对常规病例获取的RNA的分析

Fine-needle aspiration biopsy of thyroid nodules as a possible source for molecular studies: analysis of RNA obtained from routine cases.

作者信息

Mitteldorf Cristina Aparecida Troques da Silveira, de Sousa-Canavez Juliana Moreira, Massumoto Celso, da Camara-Lopes Luis Heraldo

机构信息

Laboratory of Surgical and Molecular Pathology, Hospital Sírio Libanês, rua Dona Adma Jafet 91, São Paulo, Brazil.

出版信息

Diagn Cytopathol. 2008 Dec;36(12):899-903. doi: 10.1002/dc.20939.

Abstract

Thyroid nodules are frequent in clinical practice and fine-needle aspiration biopsy (FNAB) is widely used for its evaluation, but approximately 20% of the cases are diagnosed as indeterminate for malignancy. Aspirates from thyroid nodules can be used for ancillary methods, but molecular techniques are not routinely applied to these specimens. Forty-six consecutive, routinely performed, FNAB of thyroid nodules were evaluated for the feasibility of applying RT-PCR method. RNA was extracted from 1 of 3 fresh residual samples and analyzed to determine its pureness, integrity, and concentration. Cellularity was adequate in all 46, except one, specimens analyzed, scored as 0, 1+, 2+, 3+, and 4+ in 1, 10, 14, 9, and 8 cases, respectively. Thirty-three nodules measured less than 1.5 cm. Cytological diagnosis was positive for malignancy in 3 cases, indeterminate for malignancy in 3, most probably benign follicular lesion in 7, negative for malignancy in 32, and suggestive of benign follicular lesion in 1. Good quality RNA was successfully isolated in 45/46 (97.8%) samples, with an average RNA concentration of 14 ng/microl and detection of B2M mRNA in 97.7% (44/45). There was no significant correlation between RNA concentration and nodule size or specimen cellularity. In conclusion, molecular analysis using individual, residual samples of thyroid nodules aspirates is feasible and could be employed for molecular preoperative studies in the future, adding elements for final cytological diagnosis of indeterminate cases, without altering the routine procedure.

摘要

甲状腺结节在临床实践中很常见,细针穿刺活检(FNAB)因其评估用途而被广泛应用,但约20%的病例被诊断为恶性不确定。甲状腺结节的穿刺物可用于辅助方法,但分子技术并未常规应用于这些标本。对连续46例常规进行的甲状腺结节FNAB进行评估,以确定应用逆转录聚合酶链反应(RT-PCR)方法的可行性。从3份新鲜剩余样本中的1份提取RNA,并分析其纯度、完整性和浓度。在分析的所有46份标本中,除1份外,细胞含量均充足,分别有1、10、14、9和8例标本的细胞含量评分为0、1+、2+、3+和4+。33个结节直径小于1.5 cm。细胞学诊断为恶性的有3例,恶性不确定的有3例,最可能为良性滤泡性病变的有7例,恶性阴性的有32例,提示良性滤泡性病变的有1例。45/46(97.8%)的样本成功分离出高质量RNA,平均RNA浓度为14 ng/微升,97.7%(44/45)检测到β2微球蛋白(B2M)信使核糖核酸(mRNA)。RNA浓度与结节大小或标本细胞含量之间无显著相关性。总之,使用甲状腺结节穿刺物的单个剩余样本进行分子分析是可行的,未来可用于分子术前研究,为最终细胞学诊断不确定的病例增加诊断要素,且不改变常规操作流程。

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