高通量测序数据再分析在识别结直肠癌患者ERBB2扩增中的价值  

The value of high-throughput sequencing data reanalysis in identifying ERBB2 amplification in colorectal cancer patients

作　　者：沈敏娜 张丽 陈馨宁 黄斐 白超刚 陈丽萌 彭海翔 周琰 王蓓丽 潘柏申 郭玮 SHEN Min-na;ZHANG Li;CHEN Xin-ning;HUANG Fei;BAI Chao-gang;CHEN Li-meng;PENG Hai-xiang;ZHOU Yan;WANG Bei-li;PAN Bai-shen;GUO Wei(Department of Laboratory Medicine,Zhongshan Hospital,Fudan University,Shanghai 200032,China;Department of Laboratory Medicine,Zhongshan Hospital(Xiamen Branch),Fudan University,Xiamen 361015,Fujian Province,China;Department of Laboratory Medicine,Wusong Central Hospital,Baoshan District,Shanghai 200940,China;Department of Laboratory Medicine,Shanghai Geriatric Medical Center,Shanghai 201104,China;Shanghai Dunwill Medical Technology Development Company,Shanghai 201615,China)

机构地区：[1]复旦大学附属中山医院检验科,上海200032 [2]复旦大学附属中山医院厦门医院检验科,厦门361015 [3]上海市宝山区吴淞中心医院检验科,上海200940 [4]上海市老年医学中心检验科,上海201104 [5]上海顿慧医疗科技发展有限公司,上海201615

出　　处：《复旦学报（医学版）》2024年第2期166-171,共6页Fudan University Journal of Medical Sciences

基　　金：国家自然科学基金面上项目(82172348);上海市宝山区医学重点专科项目(BSZK-2023-A18)。

摘　　要：目的评估未涵盖即ERBB2拷贝数变异(copynumbervariation,CNV)分析的高通量测序(highthroughput sequencing,HTS)数据再分析在识别结直肠癌患者ERBB2扩增中的价值。方法回顾性分析经病理活检确诊为结直肠癌并接受外周血cfDNA高通量测序样本的检测数据252例。根据经免疫组织化学(immunohistochemistry,IHC)和/或荧光原位杂交(fluorescence in situ hybridization,FISH)判断为ERBB2未扩增样本的HTS数据,计算17号染色体(Chr17)reads数占总reads数的比例范围,初步确定其为识别ERBB2扩增的阈值。根据初步阈值筛选可疑阳性样本,通过数字PCR、IHC和FISH进行验证。结果89例经IHC和/或FISH判定为ERBB2未扩增样本的Chr17 reads数与总reads数的比值为0.239±0.192(0.188~0.299)。以均数的1.25倍0.298作为识别ERBB2扩增的阈值分析其余163例样本数据,发现7例疑似阳性,比值为0.302~0.853。其中5例经IHC和/或FISH判定为阳性,6例经数字PCR验证为阳性,Chr17 reads与总reads数的比值与ERBB2/EIF2C1比值成正相关,相关性好(r2=0.909)。结论再分析未涵盖ERBB2 CNV分析的HTS数据可以初步识别结直肠癌患者的ERBB2扩增。Objective To evaluate the value of high-throughput sequencing(HTS)data reanalysis that does not include ERBB2 copy number variation(CNV)analysis,in identifying ERBB2 amplification in patients with colorectal cancer.Methods The HTS data of 252 cases of colorectal cancer diagnosed by pathological biopsy who received peripheral blood cfDNA HTS detection samples were retrospectively analyzed.According to the HTS data of ERBB2 non-amplified samples judged by immunohistochemistry(IHC)and/or fluorescence in situ hybridization(FISH),the number of chromosome 17(Chr17)reads in the total number of reads was calculated the range of the ratio was initially determined as the threshold for prompting ERBB2 amplification.Suspected positive samples were screened according to thresholds and verified by digital PCR,IHC and FISH.Results The proportion of the number of Chr17 reads accounts for the number of total reads in the 89 cases of ERBB2 non-amplified samples determined by IHC and/or FISH ranged from 0.188 to 0.299(0.239±0.192).Using 0.298(1.25 times the mean)as the threshold indicating ERBB2 amplification,the data of 163 samples were analyzed,of which 7 cases were suspected to be positive,and the ratio ranged from 0.302 to 0.853.Among them,5 cases were determined to be positive by IHC and/or FISH,and 6 cases were confirmed to be positive by digital PCR.The ratio of the number of Chr17 reads to the number of total reads was positively correlated with the ratio of ERBB2/EIF2C1,and the correlation was good(r2=0.909).Conclusion The high-throughput sequencing data that does not cover the ERBB2 CNV analysis has a certain hint value for ERBB2 amplification in patients with colorectal cancer.

关 键 词：高通量测序(HTS) 数字PCR 免疫组织化学(IHC) 荧光原位杂交(FISH) 直肠癌 

分 类 号：R446[医药卫生—诊断学]