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Clinical Chemistry 45: 777-784, 1999;
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(Clinical Chemistry. 1999;45:777-784.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Strand Displacement Amplification and Homogeneous Real-Time Detection Incorporated in a Second-Generation DNA Probe System, BDProbeTecET

Michael C. Littlea, Jeffrey Andrews, Richard Moore, Silvia Bustos, Lynda Jones, Chris Embres, Gerard Durmowicz, James Harris, Dolores Berger, Karen Yanson, Christine Rostkowski, Daretta Yursis, James Price, Thomas Fort, Adriann Walters, Matthew Collis, Oscar Llorin, Janet Wood, Frank Failing, Christian O'Keefe, Brian Scrivens, Bill Pope, Tim Hansen, Ken Marino, Keith Williams and Michael Boenisch

a Author for correspondence. Fax 410-316-3690;

Background: Amplified DNA probes provide powerful tools for the detection of infectious diseases, cancer, and genetic diseases. Commercially available amplification systems suffer from low throughput and require decontamination schemes, significant hands-on time, and specially trained laboratory staff. Our objective was to develop a DNA probe system to overcome these limitations.

Methods: We developed a DNA probe system, the BDProbeTecTMET, based on simultaneous strand displacement amplification and real-time fluorescence detection. The system uses sealed microwells to minimize the release of amplicons to the environment. To avoid the need for specially trained labor, the system uses a simple workflow with predispensed reagent devices; a programmable, expandable-spacing pipettor; and the 96-microwell format. Amplification and detection time was 1 h, with potential throughput up to 564 patient results per shift. We tested 122 total patient specimens obtained from a family practice clinic with the BD ProbeTecET and the Abbott LCx® amplified system for the detection of Chlamydia trachomatis and Neisseria gonorrhoeae.

Results: Based on reportable results, the BDProbeTecET results for both organisms were 100% sensitive and 100% specific relative to the LCx.

Conclusions: The BDProbeTecET is an easy-to-use, high-throughput, closed amplification system for the detection of nucleic acid from C. trachomatis and N. gonorrhoeae and other organisms.© 1999 American Association for Clinical Chemistry




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