基于靶向扩增子测序的快速二代测序策略在诊断髓系肿瘤中的临床应用  

Clinical application of rapid next-generation sequencing strategy based on targeted amplicon sequencing in the diagnosis of myeloid neoplasms

作　　者：解珺丹[1] 曹杨琳 张风红 姚红[1] 江艾蕊 沈宏杰[1] 岑建农[1] 吴德沛[1] 何军[1] 陈苏宁[1] Xie Jundan;Cao Yanglin;Zhang Fenghong;Yao Hong;Jiang Airui;Shen Hongjie;Cen Jiannong;Wu Depei;He Jun;Chen Suning(Diagnostic Laboratory for Hematologic Diseases,First Affiliated Hospital of Soochow University,Jiangsu Institute of Hematology,National Clinical Research Center for Hematologic Diseases,Suzhou215006,China)

机构地区：[1]苏州大学附属第一医院血液病研究室、江苏省血液研究所、国家血液系统疾病临床医学研究中心,苏州215006

出　　处：《中华检验医学杂志》2024年第11期1256-1263,共8页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金(82020108003,82170158)。

摘　　要：目的探讨基于靶向扩增子测序的快速二代测序(NGS)策略在诊断髓系肿瘤中的应用价值。方法观察性研究。收集2021年2月至2022年8月就诊于苏州大学附属第一医院的682例患者的骨髓或外周血标本,同步进行快速NGS和常规NGS检测,分别采用本地Ion Reporter软件和自主建立的生物信息学分析平台进行测序结果分析,比较2个测序平台的检测时效性,同时采用Kappa一致性检验来评估2个测序平台结果的一致性。应用快速NGS结合多重PCR和原位荧光杂交技术,在72 h内对年龄18~59岁的新诊断急性髓系白血病(AML)患者进行筛选,筛选出欧洲白血病网(ELN)2017高危组AML进行个体化诱导治疗。结果从样本接收到报告单发放,快速NGS和常规NGS检测周期时间分别为3(2,4)d和13(11,15)d,差异有统计学意义(Z=-22.636,P<0.001)。682份标本共计1507个突变位点,快速NGS共检出1499个突变位点,检出率为99.5%,检出674份结果准确,准确率为98.8%;常规NGS共检出1506个位点,检出率为99.9%,检出681份结果准确,准确率为99.9%。682份标本,181份未检出突变,501份检出突变,快速NGS漏检8份,常规NGS漏检1份。Kappa一致性检验显示,2种NGS检测结果一致性较好(Kappa=0.967,P<0.001)。通过快速诊断的286例AML患者中,筛选出ELN不良风险AML 78例,其中42例患者入组,年龄39(33,52)岁,在接受1个疗程的维奈克拉联合地西他滨诱导治疗后,78.6%(38/42)的患者获得复合完全缓解。104例其他髓系肿瘤中,快速NGS检出突变80例,检出率为76.9%,其中89.0%(215/242)的突变位点可以作为骨髓增生异常综合征、骨髓增殖性肿瘤和骨髓增生异常/骨髓增殖性肿瘤诊断分型、预后评估及靶向用药的依据。结论基于靶向扩增子测序的快速NGS与常规NGS一致性较好且缩短了诊断时间,检测灵敏度及检测范围已经满足髓系肿瘤诊断分型、预后分层及靶向治疗的需求。Objective To explore the clinical application value of rapid next-generation sequencing(NGS)strategy based on targeted amplicon sequencing in the diagnosis of myeloid neoplasms.Methods In this observational study,both rapid NGS and conventional NGS on the bone marrow or peripheral blood samples of 682 patients were prospectively performed from February 2021 to August 2022 in First Affiliated Hospital of Soochow University.The sequencing results were analyzed using the local Ion Reporter software and our lab′s self-built bioinformatics platform,respectively.The timeliness of the two sequencing platforms was compared,and the Kappa consistency test was used to evaluate the consistency between the two sequencing platforms.Patients aged between 18 and 59 years with newly diagnosed acute myeloid leukemia(AML)underwent screening by rapid NGS combining multiplex RT-PCR and in situ fluorescence hybridization technique within 72 hours,from whom high-risk patients according to European LeukemiaNet(ELN)2017 were screened for individualized induction therapy.Results In terms of timeliness,the median time from sample receipt to report issuance were 3(2,4)days and 13(11,15)days under rapid NGS and conventional NGS testing,respectively,with a statistically significant difference(Z=‒22.636,P<0.001).Among 682 specimens with a total of 1507 variants,rapid NGS detected a total of 1499 variants,with a detection rate of 99.5%and 674 cases were accurate,with an accuracy rate of 98.8%;the conventional NGS detected 1506 variants,with a detection rate of 99.9%and 681 cases were accurate,with an accuracy rate of 99.9%.In 682 specimens,there were 181 negative and 501 positive,in which 8 cases were missed under rapid NGS,and 1 case was missed under conventional NGS.The kappa value was 0.967 by Kappa consistency test,and P<0.001,suggesting good consistency and consistency between the two NGS platforms.From February 2021 to July 2022,286 patients who were rapidly diagnosed of AML contained 78 patients screened as the ELN 2017 adverse-risk

关 键 词：二代测序 突变 髓系肿瘤 诊断分型 预后分层 

分 类 号：R733[医药卫生—肿瘤]