Simultaneous Strand Displacement Amplification and Fluorescence Polarization Detection ofChlamydia trachomatisDNA
✍ Scribed by Patricia A. Spears; C.Preston Linn; Dan L. Woodard; G.Terrance Walker
- Publisher
- Elsevier Science
- Year
- 1997
- Tongue
- English
- Weight
- 144 KB
- Volume
- 247
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
which allow the use of less invasive clinical specimens Strand displacement amplification (SDA) is an isosuch as urine.
thermal DNA amplification technology that uses a re-Strand displacement amplification (SDA) is an alternastriction enzyme and polymerase. We have developed tive amplification technique that is based on a restriction a target-specific method which allows simultaneous enzyme's ability to nick a hemimodified recognition site SDA and detection in a homogeneous format. This is and the ability of the polymerase to initiate at the nick accomplished by including a detector oligodeoxyand displace the downstream strand (11-13). In general, nucleotide labeled with 5-(4,6-dichlorotriazin-2-yl)am-SDA begins with a target generation step that makes ino fluorescein in the SDA reaction. Fluorescence pocopies of the target sequence flanked by nickable restriclarization is used to monitor hybridization of the tion sites (12, 13). This is followed by exponential amplidetector probe to the amplification product as it rises fication of these modified target sequences by repeated in concentration during SDA. We have demonstrated nicking, strand displacement, and primer hybridization real-time SDA detection for the cryptic plasmid of to displaced strands (Fig. 1A). The original SDA system Chlamydia trachomatis with high sensitivity in only proceeded at 37-41ЊC and took 2 h to accomplish 10 8 -30 min.