Chehab F F, Kan Y W
Applied Biosystems, Research and Development, Foster City, CA 94404.
Proc Natl Acad Sci U S A. 1989 Dec;86(23):9178-82. doi: 10.1073/pnas.86.23.9178.
We have developed a color complementation assay that allows rapid screening of specific genomic DNA sequences. It is based on the simultaneous amplification of two or more DNA segments with fluorescent oligonucleotide primers such that the generation of a color, or combination of colors, can be visualized and used for diagnosis. Color complementation assay obviates the need for gel electrophoresis and has been applied to the detection of a large and small gene deletion, a chromosomal translocation, an infectious agent, and a single-base substitution. DNA amplification with fluorescent oligonucleotide primers has also been used to multiplex and discriminate five different amplified DNA loci simultaneously. Each primer set is conjugated to a different dye, and the fluorescence of each dye respective to its amplified DNA locus is scored on a fluorometer. This method is valuable for DNA diagnostics of genetic, acquired, and infectious diseases, as well as in DNA forensics. It also lends itself to complete automation.
我们开发了一种颜色互补检测方法，可用于快速筛选特定的基因组DNA序列。该方法基于用荧光寡核苷酸引物同时扩增两个或更多DNA片段，从而使颜色的产生或颜色组合能够被可视化并用于诊断。颜色互补检测无需进行凝胶电泳，已应用于检测大小基因缺失、染色体易位、感染因子和单碱基替换。用荧光寡核苷酸引物进行DNA扩增也已用于同时多重分析和区分五个不同的扩增DNA位点。每个引物组与一种不同的染料偶联，并且在荧光计上对每种染料相对于其扩增DNA位点的荧光进行评分。该方法对于遗传、后天和感染性疾病的DNA诊断以及DNA法医鉴定很有价值。它也适合完全自动化操作。
