胎儿染色体非整倍体检测试剂盒质量分析

Quality analysis for fetal chromosomal aneuploidy detection kit

目的使用高通量测序用外周血胎儿染色体非整倍体(T21、T18和T13)国家参考品,评价国内的胎儿染色体非整倍体21三体、18三体和13三体检测试剂盒(高通量测序法)的质量。方法依据企业各自的注册产品标准或产品技术要求,使用不同的试剂盒检测国家参考品。提取国家参考品的血浆游离DNA,通过接头连接等步骤构建文库。将文库纯化后,使用不同原理的测序仪进行检测。然后将测序数据按照试剂盒配套的软件进行分析,获得染色体非整倍体的检测结果。结果染色体正常或其他染色体异常的国家阴性参考品,结果均不是21、18和13三体。胎儿游离DNA 5%浓度的国家检测限参考品结果均是相应染色体三体;3.5%浓度的国家检测限参考品,准确检出率均不低于96%(>48/50)。70%和30%嵌合比例的国家嵌合体参考品结果均为相应染色体三体。国家微缺失微重复参考品中18号染色体微重复参考品结果均是18三体,其余参考品结果均不是21、18和13三体。结论抽检的胎儿染色体非整倍体21三体、18三体和13三体检测试剂盒均符合各自产品技术要求中阴性参考品符合率、微缺失微重复参考品符合率、嵌合体参考品符合率和检测限的要求,符合国家体外诊断试剂监督抽验的要求。

Objective To evaluate the quality of fetal trisomy 21,trisomy 18 and trisomy 13 of chromosomal aneuploidy detection kits(High⁃Throughput Sequencing)by detecting national reference materials of fetal chromosomal aneuploidy abnormality(T21,T18 and T13)in peripheral blood for next generation sequencing.Methods According to the enterprise's respective registered product standards or product technical requirements,different kits are used to detect the national reference products.The plasma cell⁃free DNA of the national reference product was extracted,and the library was constructed through the steps of linker ligation.After the library was purified,it was detected using sequencers of different principles.Then,the sequencing data was analyzed according to the software provided with the kit to obtain the detection result of chromosomal aneuploidy.Results National negative reference products with normal chromosomes or other chromosomal abnormalities were not found to be trisomy 21,18 and 13.The results of the national detection limit reference materials with a concentration of 5%of fetal cell⁃free DNA were all corresponding chromosomal trisomies;the national detection limit reference materials with a concentration of 3.5%has an accurate detection rate of not less than 96%(>48/50).The national chimerism reference materials with 70%and 30%chimerism were all corresponding chromosome trisomy.Among the national microdeletion and microduplication reference materials,the results of the chromosome 18 microduplication reference materials were all trisomy 18,and the other reference materials were not trisomy 21,18 and 13.Conclusion The sampled fetal chromosomal aneuploidy trisomy 21,trisomy 18 and trisomy 13 detection kits all meet the compliance rates of negative reference products,microdeletion and microduplication reference products,and chimera reference products in the technical requirements of their respective products and detection limit requirements.It meets the requirements of the national in vitro diagnostic reagent supervision and sampling test.