Allele identification using immobilized mismatch binding protein: detection and identification of antibiotic-resistant bacteria and determination of sheep susceptibility to scrapie
Allele identification using immobilized mismatch binding protein: detection and identification of antibiotic-resistant bacteria and determination of sheep susceptibility to scrapie
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DOI：
10.1093/nar/25.23.4825
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发表时间：
1997-12-01
影响因子：
14.9
通讯作者：
Wagner, R
中科院分区：
文献类型：
--
作者：
Debbie, P;Young, K;Wagner, R
利用固定化错配结合蛋白（IMBP）建立了一种检测和鉴定特异性等位基因的新方法。该测定涉及使用生物素标记的探针，其通过PCR扩增具有已知序列的克隆片段来制备。探针的使用避免了与IMBP测定对PCR扩增中的错误的极端敏感性相关的许多问题。该方法可用于监测PCR保真度和基因型二倍体和单倍体生物体，并已被用于区分利福平敏感和耐药菌株的结核分枝杆菌和检测和区分两个等位基因的羊朊蛋白基因参与羊瘙痒病的易感性。
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.