耳聋基因突变检测国家参考品的研制  

Development of national reference for deafness gene mutation detection

作　　者：于婷[1] 孙楠[1] 曲守方[1] 黄杰[1] YU Ting;SUN Nan;QU Shoufang;HUANG Jie(The Key Laboratory of Biotechnology Product Test Method and Its Standardization of the Ministry of Health,National Institutes for Food and Drug Control,Beijing,China,100050)

机构地区：[1]中国食品药品检定研究院卫生部生物技术产品检定方法及其标准化重点实验室

出　　处：《分子诊断与治疗杂志》2020年第1期6-10,15,共6页Journal of Molecular Diagnostics and Therapy

基　　金：国家高技术研究发展计划(863计划，2011AA02A115)

摘　　要：目的建立耳聋基因突变检测国家参考品。方法收集健康人及含20种不同耳聋基因突变位点的志愿者血清,通过Sanger测序,从中筛选出野生型和突变型的血清样本,分装制备成一套含23份样本的候选耳聋基因突变检测国家参考品。用6个厂家8个耳聋基因突变检测试剂盒对参考品进行验证。参考品的均匀性和稳定性通过联合探针锚定聚合测序法进行检测。结果各家试剂对大部分参考品的验证结果基本一致。本实验室对参考品在验证过程中新出现的突变位点进行了测序确认。参考品的均匀性和稳定性满足要求。结论通过协作验证以及均匀性、稳定性评价,该耳聋基因突变检测国家参考品可用于耳聋基因突变检测试剂盒的质量评价。Objective To establish a national reference for detection of deafness gene mutations.Methods Serum samples from healthy people and volunteers with 20 different mutation sites of deafness gene were collected.And wild type and mutant serum samples were screened out by sanger sequencing,and a set of 23 samples of candidate deafness gene mutation detection was prepared as a National Reference.8 kinds of deafness gene mutation detection kits from 6 manufacturers were used to verify the candidate reference.The homogeneity and stability of the reference were detected by the combined probe⁃anchored polymer sequencing method.Results The validation results of most reagents for most reference materials were basically the same.Some new mutation sites found in the validation process of reference were sequenced and verified in our laboratory.The homogeneity and stability of candidate reference meets the requirement.Conclusion Through the collaborative verification and the evaluation of homogeneity and stability,the national reference for deafness mutation detection can be used for the quality evaluation of deafness mutation detection kit.

关 键 词：耳聋基因突变 国家参考品 微阵列芯片法 飞行时间质谱法 联合探针锚定聚合测序法 荧光PCR法 

分 类 号：R764.43[医药卫生—耳鼻咽喉科] R440[医药卫生—临床医学]