Butler W R, Haas W H, Crawford J T
Division of AIDS, STD, and TB Laboratory Research, Centers for Disease Control and Prevention, Atlanta Georgia 30333, USA.
J Clin Microbiol. 1996 Jul;34(7):1801-3. doi: 10.1128/JCM.34.7.1801-1803.1996.
DNA fingerprints of Mycobacterium tuberculosis are produced by restriction fragment length polymorphism analysis of the insertion element IS6110. We modified a PCR-based subtyping method, mixed-linker PCR with fluorescent-labeled IS6110-specific oligonucleotides, to demonstrate rapid, automated, and unattended electrophoretic analysis. Variation in band sizing (normally occurring with fragment mobility), an artifact of lane-to-lane and gel-to-gel differences, was controlled with an internal lane standard, resulting in accurate and precise DNA sizing. By using this method, fingerprint analysis can be performed using actual fragment length rather than estimated position analysis.
结核分枝杆菌的DNA指纹图谱是通过对插入元件IS6110进行限制性片段长度多态性分析产生的。我们改进了一种基于PCR的分型方法，即使用荧光标记的IS6110特异性寡核苷酸进行混合连接子PCR，以实现快速、自动化且无人值守的电泳分析。通过内部泳道标准物来控制条带大小的变化（通常随片段迁移率出现），这是泳道间和凝胶间差异导致的假象，从而实现准确且精确的DNA大小测定。使用该方法，指纹分析可基于实际片段长度而非估计位置分析来进行。
