单细胞引物延伸预扩增分析多个基因  

Analysis of several genes using primer extension preamplification based on individual cell

作　　者：马锦琪[1] 纪亚忠[1] 盛毅[1] 赵亚南[1] 

机构地区：[1]第二军医大学附属长海医院生殖医学中心,上海200433

出　　处：《中华医学遗传学杂志》2001年第5期391-397,共7页Chinese Journal of Medical Genetics

基　　金：上海市科委重点科技项目 (9841 1 90 0 3)

摘　　要：目的　建立一种单细胞 (individual cell)水平引物延伸预扩增 (primer extensionpreamplification,PEP)方法 ,利用单个细胞同时分析多个基因。方法　采用 10 bp的随机引物先对单细胞的基因组预扩增 ,取其产物 5μl分别对 SRY、ZP3、Rh CE、Rh D、F 5个基因进行套式 PCR扩增。结果　单细胞 PEP后的产物能同时扩增 5个基因 ;对 4种已知基因型的白细胞进行诊断 ,正确率为 95 .6 %。结论PEP技术可以在单细胞水平分析 5个基因 。Objective: To analyze several genes at the same time in one individual cell using primer extension preamplification (PEP). Methods: The whole genome of individual cell was preamplified by PEP using 10 base random oligonucleotides. The aliquots (5 μl) from PEP were used to amplify SRY, ZP3, RhCE, RhD and F VIII genes by nested PCR. Results: The five genes could be amplified following PEP-PCR. Lymphocyte of four genotypes were diagnosed by PEP-PCR; correct diagnoses were obtained in 95. 6%. Conclusion At least five genes can be analyzed by PEP-PCR based on one individual cell in this laboratory. This method could be used in preimplantation genetic diagnosis and noninvasive prenatal diagnosis.

关 键 词：引物延伸预扩增 套式聚合酶链反应 胚胎种植前诊断 产前诊断 

分 类 号：R394.8[医药卫生—医学遗传学]