Allitto B A, McClatchey A I, Barnes G, Altherr M, Wasmuth J, Frischauf A M, MacDonald M E, Gusella J
Molecular Neurogenetics Laboratory, Massachusetts General Hospital, Boston.
Mol Cell Probes. 1992 Dec;6(6):513-20. doi: 10.1016/0890-8508(92)90048-3.
The Huntington's disease-linked D4S115 marker has been converted from a DNA blot assay to a more sensitive and rapid polymerase chain reaction (PCR) assay. PCR amplification of a tandem repeat at D4S115 revealed 7 allelic fragments, ranging in size from approximately 610 to 915 bp, differing in their apparent copy number of a approximately 55 bp core repeat. This repeat unit differs strikingly in sequence from the repeat units of other multi-allele markers from chromosome region 4p 16.3, arguing that the VNTR (Variable Number of Tandem Repeats) loci clustered in this region did not arise from a common ancestral sequence. The D4S115 marker can be assayed simultaneously with PCR products from D4S125, D4S95 and D4S43 on a single agarose gel, providing a rapid scan for successful amplification of these difficult-to-assay VNTRs, and for inheritance of the entire candidate Huntington's disease region. This approach should help to increase the speed, informativeness and accuracy of presymptomatic and prenatal linkage testing in this devastating disorder.
与亨廷顿舞蹈病相关的D4S115标记已从DNA印迹分析转换为更灵敏、快速的聚合酶链反应（PCR）分析。对D4S115处的串联重复序列进行PCR扩增，揭示了7个等位基因片段，大小约为610至915碱基对，其约55碱基对核心重复序列的表观拷贝数不同。该重复单元在序列上与染色体区域4p16.3的其他多等位基因标记的重复单元有显著差异，这表明聚集在该区域的VNTR（可变串联重复序列）位点并非来自共同的祖先序列。D4S115标记可与来自D4S125、D4S95和D4S43的PCR产物在同一琼脂糖凝胶上同时检测，从而快速扫描这些难以检测的VNTR的成功扩增情况，以及整个候选亨廷顿舞蹈病区域的遗传情况。这种方法应有助于提高这种毁灭性疾病的症状前和产前连锁检测的速度、信息量和准确性。
