目的:建立胎儿染色体非整倍体(21 三体、18 三体和13 三体)国家参考品,用于单核苷酸多态性(SNP)检测方法试剂盒的性能评价。方法:收集流产组织样本,提取样本的DNA。配制成10%、5% 和3.5% 等不同胎儿DNA 浓度的样本,然后进行分装。选用3 家单位对国家参考品进行协作标定。结果:对国家参考品104 例样本进行检测,其中10% 浓度阳性参考品结果均为相应染色体三体;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;5% 浓度和3.5% 浓度检测限参考品结果为相应染色体三体的检出率分别不少于87.5%(21/24)和58.0%(14/24);微缺失微重复参考品中18 号染色体微重复参考品可以被2 家单位检出18 三体,其余参考品结果均不是21、18 和13 三体。结论:研制的国家参考品,涵盖染色体非整倍体、微缺失微重复和正常样本,可以用于相关试剂盒的性能评价及上市后的监督管理工作。
Objective: To establish the national reference materials for the fetal chromosomal aneuploidies(trisomy 21,trisomy 18,and trisomy 13),and evaluate the performance of detection kits baseed on single nucleotide polymorphism(SNP)method. Methods: Tissue samples of abortion were collected and DNA was extracted. Samples with different fetal DNA concentrations of 10%,5% and 3.5% were prepared and then repackaged. 3 companies participated in collaborative calibration to evaluate the national reference material. Results: 104 samples of the national reference materials were tested,among which the positive reference samples with 10% concentration were all corresponding trisomy. For samples with normal or other chromosomal abnormalities,the results were not trisomy 21,18 or 13. The detection rate of detection limit reference samples with 5% and 3.5% concentration was not less than 87.5%(21/24)and 58.0%(14/24),respectively. Among the reference samples with microdeletion and microduplication,the samples with microduplication of chromosome 18 were trisomy 18 by 2 companies,while the results of the other samples were not trisomy 21,18 or 13. Conclusion: The developed national reference materials cover chromosome aneuploidy,microdeletion and microduplication and normal samples,and can be used for the performance evaluation of the relevant kits and the supervision and management after it comes into the market.
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