%0 Journal Article
%T Development of 3-enzyme pyrosequencing system and its application in rapid diagnosis of Down’s syndrome<br>三酶焦测序体系的建立及其在唐氏综合征快速诊断中的应用
%A 刘夕群
%A 朱术会
%A 邹秉杰 马寅姣
%A 周国华
%J 遗传
%D 2010
%I 
%X To avoid sequencing error resulting from use of apyrase in conventional 4- enzyme pyrosequencing system, a non-apyrase 3-enzyme pyrosequencing system with a better performance of quantitative analysis was established. The method is to immobilize biotinylated DNA template, ATP sulfurylase and luciferase on streptavidin-coated magnetic beads for pyrosequencing. After pyrosequencing, ATP produced from the pyrosequencing reaction and excess dNTPs were removed by magnetic separation technique; another dNTP was then dispensed for sequencing reaction, and the components interfering with the next circle of pyrosequencing reaction were removed by the same way, achieving the circular sequenc-ing. This new system can accurately measure base sequences of a target DNA template, and also can quantitatively deter-mine the relative ratio of two alleles. The allele ratios in two SNPs (rs1042917 and rs4818219) having a higher heterozygote rate on chromosome 21 were successfully detected for 16 normal samples and 8 clinical samples from Down’s syndrome patients. The results can accurately demonstrate whether or not the target sample has equal copies of chromosome 21 from mother and father. This paper established a non-apyrase 3-enzyme pyrosequencing method, which owns a good perform-ance of quantitative analysis. The method is especially suitable to allelic quantification of an SNP, enabling the rapid diag-nosis of Down’s syndrome by analyzing allele ratio of SNPs on chromosome 21.
%K 腺苷双磷酸酶
%K 三酶焦测序体系
%K 唐氏综合征
%K 单核苷酸序列多态性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=3E23F50E071DF18E21B2F5AEE4F1FA5E&aid=3C71F9A0192D9B612F23D973791D207E&yid=140ECF96957D60B2&vid=9971A5E270697F23&iid=94C357A881DFC066&sid=5A751AE9FA58A3FB&eid=C81D738643975BB0&journal_id=0253-9772&journal_name=遗传&referenced_num=0&reference_num=11