Debbie P, Young K, Pooler L, Lamp C, Marietta P, Wagner R
Gene Check Inc., Suite 106, 1629 Blue Spruce Drive, Fort Collins, CO 80524, USA.
Nucleic Acids Res. 1997 Dec 1;25(23):4825-29. doi: 10.1093/nar/25.23.4825.
A novel method for detection and identification of specific alleles has been developed utilizing immobilized mismatch binding protein (IMBP). The assay involves the use of biotin-labeled probes, which are prepared by PCR amplification of cloned fragments with known sequence. The use of probes avoids many of the problems associated with the extreme sensitivity of IMBP assays to errors in PCR amplification. The method can be used to monitor PCR fidelity and to genotype both diploid and haploid organisms and has been used to distinguish rifampicin-sensitive and -resistant strains of Mycobacterium tuberculosis and to detect and distinguish two alleles of the sheep prion protein gene involved in susceptibility to scrapie.
利用固定化错配结合蛋白（IMBP）开发了一种检测和鉴定特定等位基因的新方法。该检测方法涉及使用生物素标记的探针，这些探针是通过对已知序列的克隆片段进行PCR扩增制备的。探针的使用避免了许多与IMBP检测对PCR扩增错误的极端敏感性相关的问题。该方法可用于监测PCR保真度，并对二倍体和单倍体生物进行基因分型，已用于区分结核分枝杆菌的利福平敏感和耐药菌株，以及检测和区分绵羊朊病毒蛋白基因中与痒病易感性相关的两个等位基因。
