Detection of Escherichia coli rRNA using target amplification and time-resolved fluorescence detection
The development of technology to increase the sensitivity and speed of detection of bacterial pathogens in samples is important for diagnosis and monitoring of illness. We have developed a sensitive and rapid method for the detection of bacteria, using Escherichia coli as a model, which combines tra...
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| Veröffentlicht in: | Molecular and cellular probes 1991-12, Vol.5 (6), p.467-472 |
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| container_title | Molecular and cellular probes |
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| creator | Bush, Charlene E. Vanden Brink, Kurt M. Sherman, David G. Richard Peterson, W. Beninsig, Laura A. Godsey, James H. |
| description | The development of technology to increase the sensitivity and speed of detection of bacterial pathogens in samples is important for diagnosis and monitoring of illness. We have developed a sensitive and rapid method for the detection of bacteria, using Escherichia coli as a model, which combines transcription-based target amplification with a bead-based sandwich hybridization assay using rare earth metal chelate labelled probes and time-resolved fluorescence detection. Using these methods as little as 100 copies (0·00016 attomoles) of purified native Escherichia coli rRNA or just one bacterial cell in a spiked sample could be detected. These results demonstrate that amplification of rRNA by transcription-based amplification and detection by time-resolved fluorescence provide a sensitive technology for the direct detection of micro-organisms without the requirement for prior cultivation. |
| doi_str_mv | 10.1016/S0890-8508(05)80019-2 |
| format | Article |
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We have developed a sensitive and rapid method for the detection of bacteria, using Escherichia coli as a model, which combines transcription-based target amplification with a bead-based sandwich hybridization assay using rare earth metal chelate labelled probes and time-resolved fluorescence detection. Using these methods as little as 100 copies (0·00016 attomoles) of purified native Escherichia coli rRNA or just one bacterial cell in a spiked sample could be detected. 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We have developed a sensitive and rapid method for the detection of bacteria, using Escherichia coli as a model, which combines transcription-based target amplification with a bead-based sandwich hybridization assay using rare earth metal chelate labelled probes and time-resolved fluorescence detection. Using these methods as little as 100 copies (0·00016 attomoles) of purified native Escherichia coli rRNA or just one bacterial cell in a spiked sample could be detected. These results demonstrate that amplification of rRNA by transcription-based amplification and detection by time-resolved fluorescence provide a sensitive technology for the direct detection of micro-organisms without the requirement for prior cultivation.</abstract><cop>Kidlington</cop><pub>Elsevier Ltd</pub><pmid>1723491</pmid><doi>10.1016/S0890-8508(05)80019-2</doi><tpages>6</tpages></addata></record> |
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| identifier | ISSN: 0890-8508 |
| ispartof | Molecular and cellular probes, 1991-12, Vol.5 (6), p.467-472 |
| issn | 0890-8508 1096-1194 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_72629324 |
| source | MEDLINE; Elsevier ScienceDirect Journals |
| subjects | Bacteriological methods and techniques used in bacteriology Bacteriology Base Sequence Biological and medical sciences Escherichia coli Escherichia coli - isolation & purification Fluorescent Dyes Fundamental and applied biological sciences. Psychology Humans Metals, Rare Earth Microbiology Microspheres Molecular Sequence Data Nucleic Acid Amplification Techniques Oligonucleotide Probes Polystyrenes RNA, Bacterial - genetics RNA, Bacterial - isolation & purification RNA, Ribosomal - genetics RNA, Ribosomal - isolation & purification Sensitivity and Specificity time-resolved fluorescence detection transcription-based amplification |
| title | Detection of Escherichia coli rRNA using target amplification and time-resolved fluorescence detection |
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