Haas W H, Butler W R, Woodley C L, Crawford J T
National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333.
J Clin Microbiol. 1993 May;31(5):1293-8. doi: 10.1128/jcm.31.5.1293-1298.1993.
Rapid recognition of multidrug-resistant strains of Mycobacterium tuberculosis is a desirable goal for treatment of patients and protection of health care workers. DNA fingerprints produced with the insertion sequence IS6110 generate restriction fragment length polymorphism (RFLP) patterns that reliably identify M. tuberculosis complex strains. This report describes a rapid technique for RFLP typing using the polymerase chain reaction. The method uses one primer specific for IS6110 and a second primer complementary to a linker ligated to the restricted genomic DNA. In one strand the linker contains uracil in place of thymidine, and specific amplification is obtained by elimination of this strand with uracil N-glycosylase. Mixed-linker fingerprinting clearly differentiated multidrug-resistant isolates from 12 outbreaks and unambiguously assigned them to 26 RFLP groups.
快速识别结核分枝杆菌的多重耐药菌株是治疗患者和保护医护人员的一个理想目标。用插入序列IS6110产生的DNA指纹图谱可生成限制性片段长度多态性（RFLP）模式，从而可靠地鉴定结核分枝杆菌复合群菌株。本报告描述了一种使用聚合酶链反应进行RFLP分型的快速技术。该方法使用一种对IS6110特异的引物和另一种与连接到经酶切的基因组DNA上的接头互补的引物。在一条链中，接头含有尿嘧啶而不是胸腺嘧啶，通过用尿嘧啶N-糖基化酶去除这条链来实现特异性扩增。混合接头指纹图谱能清晰地区分12起暴发中的多重耐药分离株，并明确地将它们归为26个RFLP组。
