Walker G T, Nadeau J G, Spears P A, Schram J L, Nycz C M, Shank D D
Becton Dickinson Research Center, Research Triangle Park, NC 27709-2016.
Nucleic Acids Res. 1994 Jul 11;22(13):2670-7. doi: 10.1093/nar/22.13.2670.
Strand Displacement Amplification (SDA) is an isothermal, in vitro method of amplifying a DNA target sequence prior to detection [Walker et al (1992) Nucleic Acids Res., 20, 1691-1693]. Here we describe a multiplex form of SDA that allows two target sequences and an internal amplification control to be co-amplified by a single pair of primers after common priming sequences are spontaneously appended to the ends of target fragments. Multiplex SDA operates at a single temperature, under the same simple protocol previously developed for single-target SDA. We applied multiplex SDA to co-amplification of a target sequence (IS6110) that is specific to members of the Mycobacterium tuberculosis-complex and a target (16S ribosomal gene) that is common to most clinically relevant species of mycobacteria. Both targets are amplified 10(8)-fold during a 2 hour, single temperature incubation. The relative sensitivity of the system was evaluated across a number of clinically relevant mycobacteria and checked for crossreactivity against organisms that are closely related to mycobacteria.
链置换扩增（SDA）是一种等温体外方法，用于在检测前扩增DNA靶序列[Walker等人（1992年），《核酸研究》，20，1691 - 1693]。在此，我们描述了一种多重SDA形式，在靶片段末端自发添加共同引物序列后，该方法允许通过一对引物共同扩增两个靶序列和一个内部扩增对照。多重SDA在单一温度下运行，遵循先前为单靶SDA开发的相同简单方案。我们将多重SDA应用于共同扩增结核分枝杆菌复合群成员特有的靶序列（IS6110）和大多数临床相关分枝杆菌物种共有的靶标（16S核糖体基因）。在2小时的单一温度孵育过程中，两个靶标均扩增了10^8倍。在多种临床相关分枝杆菌中评估了该系统的相对灵敏度，并检查了其对与分枝杆菌密切相关的生物体的交叉反应性。
