Lo Y M, Patel P, Newton C R, Markham A F, Fleming K A, Wainscoat J S
Nuffield Department of Pathology and Bacteriology, John Radcliffe Hospital, Oxford, UK.
Nucleic Acids Res. 1991 Jul 11;19(13):3561-7. doi: 10.1093/nar/19.13.3561.
We have developed a novel double Amplification Refractory Mutation System (double ARMS) using a highly polymorphic region 5' to the human delta-globin gene as a model system. The double ARMS approach involves using two allele-specific ARMS primers simultaneously during DNA amplification by the polymerase chain reaction (PCR). The resulting system is highly sensitive and more specific than single ARMS. In addition, this approach enables the elucidation of the relationship of polymorphic sites on the same chromosome and thus allows the direct determination of haplotypes. We have also demonstrated that this system can be used in conjunction with inverse PCR, the resulting double ARMS inverse PCR (DARMSI-PCR) may allow haplotype determination on polymorphic sites which are separated further apart than the length limit imposed by PCR. The double ARMS approach has numerous other applications in molecular biology including HLA typing, virology, forensic pathology and the investigation of the phenomenon of chimerism following bone marrow transplantation.
我们利用人类δ-珠蛋白基因5'端的一个高度多态性区域作为模型系统，开发了一种新型的双重扩增不应突变系统（双重ARMS）。双重ARMS方法涉及在通过聚合酶链反应（PCR）进行DNA扩增时同时使用两种等位基因特异性ARMS引物。所得系统比单一ARMS具有更高的灵敏度和特异性。此外，这种方法能够阐明同一染色体上多态性位点的关系，从而直接确定单倍型。我们还证明了该系统可与反向PCR结合使用，由此产生的双重ARMS反向PCR（DARMSI-PCR）可能允许在比PCR施加的长度限制分隔得更远的多态性位点上确定单倍型。双重ARMS方法在分子生物学中有许多其他应用，包括HLA分型、病毒学、法医病理学以及骨髓移植后嵌合体现象的研究。
