ENZYMATIC AMPLIFICATION OF BETA-GLOBIN GENOMIC SEQUENCES AND RESTRICTION SITE ANALYSIS FOR DIAGNOSIS OF SICKLE-CELL ANEMIA
ENZYMATIC AMPLIFICATION OF BETA-GLOBIN GENOMIC SEQUENCES AND RESTRICTION SITE ANALYSIS FOR DIAGNOSIS OF SICKLE-CELL ANEMIA
复制标题
DOI：
10.1126/science.2999980
复制
发表时间：
1985-01-01
期刊：
影响因子：
56.9
通讯作者：
ARNHEIM, N
中科院分区：
文献类型：
--
作者：
SAIKI, RK;SCHARF, S;ARNHEIM, N
采用两种新方法建立了镰状细胞性贫血的快速、高灵敏度产前诊断试验。第一种涉及引物介导的基因组DNA中特定β-珠蛋白靶序列的酶扩增，导致靶DNA拷贝呈指数增长（22万倍）。在第二种技术中，通过限制性内切酶酶切末端标记的寡核苷酸探针来确定β a和β小基因的存在，探针与扩增的β-珠蛋白序列杂交在溶液中。在含有显著少于1微克基因组DNA的样品中，可以在不到1天的时间内确定β-珠蛋白基因型。
Two new methods were used to establish a rapid and highly sensitive prenatal diagnostic test for sickle cell anemia. The first involves the primer-mediated enzymatic amplification of specific β-globin target sequences in genomic DNA, resulting in the exponential increase (220,000 times) of target DNA copies. In the second technique, the presence of the βAand βSalleles is determined by restriction endonuclease digestion of an end-labeled oligonucleotide probe hybridized in solution to the amplified β-globin sequences. The β-globin genotype can be determined in less than 1 day on samples containing significantly less than 1 microgram of genomic DNA.