Clinical Chemistry
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Clinical Chemistry 42: 9-13, 1996;
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Clinical Chemistry, Vol 42, 9-13, Copyright © 1996 by American Association for Clinical Chemistry

Strand displacement amplification (SDA) and transient-state fluorescence polarization detection of Mycobacterium tuberculosis DNA

GT Walker, JG Nadeau, CP Linn, RF Devlin and WB Dandliker
Becton Dickinson Research Center, Research Triangle Park, NC 27709- 2016, USA. walker@bdrc.bd.com

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.


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