Variation Scanning with Mismatch Repair Detection MRD)

描述（由申请人提供）：一种被广泛考虑的鉴定易感等位基因的方法是通过这些等位基因之间的连锁不平衡（LD）和分布在所有或大多数种族人群中的一组普遍的相对频繁的snp。然而，当导致疾病易感性的变异等位基因不常见或仅针对特定人群时，这种方法可能是不现实的。在这种情况下，确定易感等位基因可能需要在患者和对照组之间进行全面的序列比较。对于未知变异的高通量扫描技术缺乏。错配修复检测（MRD）先前已经描述过，它利用细菌错配修复系统在体内检测人类DNA样本中的序列变异。错配修复检测（MRD）潜在的高通量扫描可用于全面比较患者和对照组之间的序列。MRD以前已经描述过，它利用细菌错配修复系统在体内检测人类DNA样本中的序列变异。许多片段可以被引入到一个特定的细菌菌株突变分选器（MS）中，该分选器被设计成将这些片段分为两个池：携带变异的和不携带变异的。DNA变异检测的问题然后减少到鉴定两个池的片段内容的问题，这可以使用微阵列杂交来完成。在第一阶段，我们的目标是展示MRD同时扫描500个外显子变化的能力，使用通用芯片作为最终读数。

DESCRIPTION (provided by applicant): One widely considered approach to identify susceptibility alleles is through linkage disequilibrium (LD) between these alleles and a universal set of relatively frequent SNPs distributed in all or most ethnic populations. This approach, however, may be unrealistic when variant alleles responsible for disease susceptibility are infrequent or are specific to a particular population. In such cases, identifying susceptibility alleles may require comprehensive sequence comparison between patients and controls. There is a paucity of techniques for high throughput scanning for unknown variations. Mismatch Repair Detection (MRD) has been described previously and it utilizes a bacterial mismatch repair system in vivo to detect sequence variants in human DNA samples. Mismatch Repair Detection (MRD)'s potential for high throughput scanning can be used to comprehensively compare sequences between patients and controls. MRD has been described previously and it utilizes a bacterial mismatch repair system in vivo to detect sequence variants in human DNA samples. Many fragments can be introduced into a specific bacterial strain Mutation Sorter (MS) that is engineered to sort these fragments to two pools: those carrying variations and those that do not. The problem of DNA variation detection is then reduced to the problem of identification of the fragment content of the two pools, and that can be done using a microarray hybridization. In phase I of this grant we aim to demonstrate the ability of MRD to scan for variations 500 exons simultaneously using a generic chip as the final readout.