Waldenström E, Lagerkvist A, Dahlman T, Westermark K, Landegren U
Beijer Laboratory, Department of Medical Genetics, Uppsala Biomedical Center, Sweden.
Genomics. 1996 Nov 1;37(3):303-9. doi: 10.1006/geno.1996.0564.
We have applied a solid support for parallel handling and direct loading of sequencing reactions--manifold sequencing--to analyze the coding sequence for the deficient copper transporting P-type ATPase in 24 families with Wilson disease. At least 100 different amplification reactions could be handled in parallel, with a minimal turnaround time of 12 h from isolated genomic DNA to identification of the mutations. Sixteen different mutations were found, accounting for 92% of the mutant genes. Ten of these mutations have not been previously described. Eleven were observed only in single families. Mutation His1069Gln, previously identified as the most prevalent mutation in Northern Europe, was found in one-third of the Northern European chromosomes in our material. Four patients were homozygous for this mutation, and three were homozygous for Thr977Met. The method allowed us to establish the diagnosis of Wilson disease in 24 h in a patient with acute hepatic failure.
我们应用了一种用于平行处理和直接加载测序反应的固相支持物——多重测序，来分析24个威尔逊病家族中缺陷型铜转运P型ATP酶的编码序列。至少100种不同的扩增反应可以并行处理，从分离的基因组DNA到鉴定突变的最短周转时间为12小时。发现了16种不同的突变，占突变基因的92%。其中10种突变以前未曾描述过。11种仅在单个家族中观察到。先前在北欧被确定为最常见突变的His1069Gln突变，在我们材料中三分之一的北欧染色体中被发现。4名患者该突变呈纯合状态，3名患者Thr977Met呈纯合状态。该方法使我们能够在24小时内对一名急性肝衰竭患者做出威尔逊病的诊断。
