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(R) 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.
%0 Journal Article
%1 little_strand_1999
%A Little, M C
%A Andrews, J
%A Moore, R
%A Bustos, S
%A Jones, L
%A Embres, C
%A Durmowicz, G
%A Harris, J
%A Berger, D
%A Yanson, K
%A Rostkowski, C
%A Yursis, D
%A Price, J
%A Fort, T
%A Walters, A
%A Collis, M
%A Llorin, O
%A Wood, J
%A Failing, F
%A O'Keefe, C
%A Scrivens, B
%A Pope, B
%A Hansen, T
%A Marino, K
%A Williams, K
%D 1999
%J Clinical Chemistry
%K Amplification, Bacterial, Chlamydia Diagnostic, Fluorescence, Gene Humans, Kits, Neisseria Probes, Reagent Sensitivity Specificity and gonorrhoeae, trachomatis, {DNA,} {DNA}
%N 6 Pt 1
%P 777--784
%T Strand displacement amplification and homogeneous real-time detection incorporated in a second-generation DNA probe system, BDProbeTecET
%U http://www.ncbi.nlm.nih.gov/pubmed/10351985
%V 45
%X 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(R) 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.
@article{little_strand_1999,
abstract = {{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(R)} 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.},
added-at = {2011-03-11T10:05:34.000+0100},
author = {Little, M C and Andrews, J and Moore, R and Bustos, S and Jones, L and Embres, C and Durmowicz, G and Harris, J and Berger, D and Yanson, K and Rostkowski, C and Yursis, D and Price, J and Fort, T and Walters, A and Collis, M and Llorin, O and Wood, J and Failing, F and {O'Keefe}, C and Scrivens, B and Pope, B and Hansen, T and Marino, K and Williams, K},
biburl = {https://www.bibsonomy.org/bibtex/208bf4bf729485ba355fbef6405fe286a/jelias},
interhash = {317693e3ea83ae8f4b7888d2ff47246f},
intrahash = {08bf4bf729485ba355fbef6405fe286a},
issn = {0009-9147},
journal = {Clinical Chemistry},
keywords = {Amplification, Bacterial, Chlamydia Diagnostic, Fluorescence, Gene Humans, Kits, Neisseria Probes, Reagent Sensitivity Specificity and gonorrhoeae, trachomatis, {DNA,} {DNA}},
month = jun,
note = {{PMID:} 10351985},
number = {6 Pt 1},
pages = {777--784},
timestamp = {2011-03-11T10:05:41.000+0100},
title = {Strand displacement amplification and homogeneous real-time detection incorporated in a second-generation {DNA} probe system, {BDProbeTecET}},
url = {http://www.ncbi.nlm.nih.gov/pubmed/10351985},
volume = 45,
year = 1999
}