两种CAG动态突变（亨廷顿舞蹈症和1型脊髓小脑共济失调）的印迹PCR产物的化学发光检测（CB-PCR）

Castellví-Bel S, Matilla T, Banchs M I, Kruyer H, Corral J, Milà M, Estivill X

Genetics Service, Hospital Clínic i Provincial, Barcelona, Catalunya, Spain.

J Med Genet. 1994 Aug;31(8):654-5. doi: 10.1136/jmg.31.8.654.

We have used a non-isotopic PCR assay based on the chemiluminescent detection of blotted PCR products (CB-PCR) for two dynamic mutation diseases (Huntington's disease and spinocerebellar ataxia type 1). This gives an accurate sizing of alleles and permits a rapid analysis of at risk persons. The system involves PCR of the samples, separation of alleles on polyacrylamide gels, Southern blotting, and hybridisation with specific primers 3' labelled with fluorescein (F1)-dUTP as probes. CB-PCR retains the isotopic sensitivity for accurate allele determination, avoids isotopic manipulation, and provides the advantages of safety, long term storage of probes, and recycling of hybridisation solutions.

我们已将基于印迹PCR产物化学发光检测的非同位素PCR检测法（CB-PCR）用于两种动态突变疾病（亨廷顿舞蹈病和1型脊髓小脑共济失调）。这能准确测定等位基因大小，并可快速分析高危人群。该系统包括样本的PCR、在聚丙烯酰胺凝胶上分离等位基因、Southern印迹以及用3'端标记有荧光素（F1）-dUTP的特异性引物作为探针进行杂交。CB-PCR保留了用于准确确定等位基因的同位素敏感性，避免了同位素操作，并具有安全性高、探针可长期保存以及杂交溶液可循环利用等优点。