Novel, ultrasensitive, Q-beta replicase-amplified hybridization assay for detection of Chlamydia trachomatis
| Autor: | S. Popoff, J. Smith, W. King, G. Serpe, J Liu, D. M. Olive, W. J. O'brien, G Radcliffe, J. S. Shah |
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| Rok vydání: | 1994 |
| Předmět: |
Microbiology (medical)
Molecular Sequence Data RNA-dependent RNA polymerase Chlamydia trachomatis medicine.disease_cause Sensitivity and Specificity Bacteriophage Nucleic acid thermodynamics chemistry.chemical_compound medicine Humans Propidium iodide Base Sequence biology Gene Amplification Q beta Replicase Nucleic Acid Hybridization RNA biology.organism_classification Molecular biology RNA Bacterial chemistry RNA Ribosomal Nucleic acid Research Article |
| Zdroj: | Journal of Clinical Microbiology. 32:2718-2724 |
| ISSN: | 1098-660X 0095-1137 |
| DOI: | 10.1128/jcm.32.11.2718-2724.1994 |
| Popis: | A sensitive, nonisotopic hybridization assay termed "dual capture" is described. The assay rapidly and specifically detects very low levels of target nucleic acids and organisms. The assay is based on the principles of sandwich hybridization, reversible target capture, and Q-Beta replicase amplification. The assay can be completed in less than 4 h, and in the described model format, it detects Chlamydia trachomatis rRNA or rDNA. Up to 96 samples can be analyzed simultaneously. The assay employs two types of probes: a test-specific capture probe, which mediates the cycling of the target probe complex on and off derivatized magnetic beads, and a replicatable RNA detector molecule containing a sequence complementary to and adjacent to the capture probe site on the target. Following reversible target capture, detection of the signal is accomplished by replication of the detector molecule by Q-Beta replicase in the presence of propidium iodide. A specific assay signal can be detected from as few as 1,000 molecules above the background. In a limited study of 94 urogenital samples the assay detected five of the six culture-positive samples and did not detect the C. trachomatis target in 85 of the 88 culture-negative samples. |
| Databáze: | OpenAIRE |
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