Walker G T, Nadeau J G, Linn C P, Devlin R F, Dandliker W B
Becton Dickinson Research Center, Research Triangle Park, NC 27709-2016, USA.
Clin Chem. 1996 Jan;42(1):9-13.
Strand displacement amplification (SDA) is an isothermal, in vitro method of amplifying a DNA sequence for diagnostic purposes. We have combined SDA with fluorescence polarization detection in a closed, homogeneous format. A fluorescently labeled oligodeoxynucleotide detector probe hybridizes to the amplification product that increases in concentration during SDA. The single- to double-stranded conversion of the probe is accompanied by an increase in fluorescence polarization values, which can be measured in real-time without physical manipulation of the sample. The probe was labeled with the near-infrared dye La Jolla Blue, and fluorescence polarization was measured on a transient-state fluorometer. We have applied this homogeneous SDA/detection system to a target DNA sequence specific for Mycobacterium tuberculosis DNA.
链置换扩增（SDA）是一种用于诊断目的的等温体外DNA序列扩增方法。我们已将SDA与荧光偏振检测相结合，形成一种封闭的均相检测形式。一种荧光标记的寡脱氧核苷酸检测探针与在SDA过程中浓度增加的扩增产物杂交。探针从单链到双链的转变伴随着荧光偏振值的增加，可在不对样品进行物理操作的情况下实时测量。该探针用近红外染料拉霍亚蓝标记，并在瞬态荧光计上测量荧光偏振。我们已将这种均相SDA/检测系统应用于结核分枝杆菌DNA特有的靶DNA序列。
