cDNA-Based Mutation Screening Using a Combination of High-Resolution Melting Curve and Fragment Analysis Facilitates Efficient CCR4 Mutation Analysis in Adult T-Cell Leukemia/Lymphoma
cDNA-Based Mutation Screening Using a Combination of High-Resolution Melting Curve and Fragment Analysis Facilitates Efficient CCR4 Mutation Analysis in Adult T-Cell Leukemia/Lymphoma
复制标题
结合高分辨率熔解曲线和片段分析进行基于 cDNA 的突变筛选，有助于对成人 T 细胞白血病/淋巴瘤进行有效的 CCR4 突变分析
DOI：
10.1093/ajcp/aqaa037
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发表时间：
2020
影响因子：
3.5
通讯作者：
Mitsumasa Watanabe
中科院分区：
文献类型：
--
作者：
Shumpei Mizuta;Noriko Yamane;Saya Mononobe;Takao Komai;Yusuke Koba;Takahito Kawata;Naoya Ukyo;Akira Tamekane;Mitsumasa Watanabe
目的C-C趋化因子受体4(CCR4)蛋白在成人T细胞白血病/淋巴瘤(ATLL)的肿瘤细胞上表达。鉴于CCR4基因突变状态与临床预后和莫古珠单抗疗效等重要临床信息相关，我们旨在建立一种适用于临床实验室检测的筛查方法。方法对34例ATLL患者进行CCR4突变分析、高分辨熔融(HRM)分析、片段分析和直接测序。对38例人类T细胞白血病病毒1型(HTLV-1)无症状携带者进行CCR4突变筛查。结果34份临床标本直接测序突变分析，在4份基因组DNA和7份cDNA样本中发现CCR4突变，并鉴定出2个新突变。在HTLV-1无症状携带者中未检测到CCR4突变。结论HRM和片段分析相结合的方法筛查CCR4突变是一种简单实用的方法，有助于更好地决策治疗策略，为临床医生提供快速的CCR4突变状态。
ObjectivesC-C chemokine receptor type 4 (CCR4) proteins are expressed on the neoplastic cells of adult T-cell leukemia/lymphoma (ATLL). As the mutation status ofCCR4gene is reported to correlate with significant clinical information such as prognosis and response to mogamulizumab, we aimed to establish a screening method that is suitable for clinical laboratory tests.MethodsIn 34 patients with ATLL,CCR4mutation analysis, high-resolution melting (HRM) analysis, fragment analysis, and direct sequencing were performed using both genomic DNA and complementary DNA (cDNA). Furthermore, 38 cases of asymptomatic carriers of human T-cell leukemia virus type 1 (HTLV-1) were screened forCCR4mutation.ResultsMutation analysis by direct sequencing of 34 ATLL clinical samples detectedCCR4mutation in four genomic DNA samples and seven cDNA samples, and two novel mutations were identified. AllCCR4mutations detected by direct sequencing were positive for HRM analysis and/or fragment analysis.CCR4mutation was not detected in the asymptomatic carriers of HTLV-1.ConclusionsCCR4mutation screening by a combination of HRM and fragment analysis using cDNA is a simple and practical method, and it will contribute to better decision making for a therapeutic strategy, providing a rapidCCR4mutational status to clinicians.