Vuorio A F, Paulin L, Turtola H, Kontula K
Institute of Biotechnology, University of Helsinki, Finland.
Mol Cell Probes. 1997 Feb;11(1):65-70. doi: 10.1006/mcpr.1996.0078.
We evaluated the feasibility of methods based on the polymerase chain reaction (PCR) and non-automated or automated gel electrophoresis to detect clinically important DNA deletions in pooled DNA and blood samples. Two common low density lipoprotein (LDL) receptor mutations causing familial hypercholesterolaemia (FH) in the Finnish population were easily identified in pools corresponding to 20 individuals. One of these mutations (FH-North Karelia) deletes seven nucleotides from exon 6 of the LDL receptor gene. PCR amplification of DNA samples from the heterozygous patients with the FH-North Karelia gene results in the formation of DNA heteroduplexes, which markedly improves mutation detection. These studies show the applicability of semi-automated PCR techniques in the screening of DNA deletions and demonstrate the clinical diagnostic usefulness of heteroduplex formation.
我们评估了基于聚合酶链反应（PCR）以及非自动化或自动化凝胶电泳的方法在检测混合DNA和血液样本中临床重要DNA缺失方面的可行性。在芬兰人群中导致家族性高胆固醇血症（FH）的两种常见的低密度脂蛋白（LDL）受体突变，在对应20个人的样本池中很容易被识别出来。其中一种突变（FH - 北卡累利阿）从LDL受体基因的外显子6中缺失了七个核苷酸。对携带FH - 北卡累利阿基因的杂合患者的DNA样本进行PCR扩增会导致DNA异源双链体的形成，这显著提高了突变检测的效率。这些研究表明半自动PCR技术在DNA缺失筛查中的适用性，并证明了异源双链体形成在临床诊断中的有用性。
