PCR: Application of nested PCR to detection of mycoplasmas
PCR: Application of nested PCR to detection of mycoplasmas
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PCR：巢式PCR在支原体检测中的应用
DOI：
10.1016/b978-012583806-1/50119-x
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发表时间：
1996
影响因子：
5.6
通讯作者：
R. Harasawa
中科院分区：
文献类型：
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作者：
R. Harasawa
聚合酶链式反应 (PCR) 最流行的改进之一，称为巢式 PCR（Mullis 和 Faloona，1987），是使用巢式引物对进行体外 DNA 扩增。在巢式 PCR 的典型方案中，第一轮 PCR 使用一对外部引物进行。将少量第一轮PCR产物转移至新的反应管中，使用一对内引物进行第二轮PCR。巢式 PCR 是目前检测细胞培养物或临床材料中支原体最灵敏的方法，因为可以检测到单个拷贝的靶序列（Harasawa 等，1993a，b）。巢式PCR的另一个优点是第二轮PCR可以确认第一轮PCR的特异性。本章推荐的嵌套引物对由 16S–23S rRNA 基因间间隔区序列组成（图 1）。
One of the most popular modifications of polymerase chain reaction (PCR), known as nested PCR (Mullis and Faloona, 1987), is an in vitro DNA amplification employing nested primer pairs. In a typical protocol for the nested PCR, a first-round PCR is performed with a pair of outer primers. A small amount of the first-round PCR product is transferred to a fresh reaction tube for a second-round PCR by using a pair of inner primers. The nested PCR is currently the most sensitive means of detecting the mycoplasmas in cell cultures or in clinical materials because a single copy of target sequence can be detected (Harasawa et al., 1993a, b). Another advantage of the nested PCR is that the second-round PCR serves to confirm the specificity of the first-round PCR. The nested primer pairs recommended in this chapter consist of sequences of the 16S–23S rRNA intergenic spacer regions (Fig. 1).