Rapid and sensitive PCR-based detection and differentiation of aetiologic agents of human granulocytotropic and monocytotropic ehrlichiosis
| Autor: | Frederick K. Chu |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
DNA
Bacterial Ehrlichia Polymerase Chain Reaction Sensitivity and Specificity Monocytes law.invention chemistry.chemical_compound law RNA Ribosomal 16S Animals Humans Ehrlichia chaffeensis Magnesium Horses Molecular Biology Polymerase chain reaction DNA Primers Whole blood biology Ehrlichiosis Nucleic Acid Hybridization Cell Biology biology.organism_classification 16S ribosomal RNA Molecular biology chemistry Ehrlichiosis (canine) Human Ehrlichioses Nucleic acid DNA Probes DNA Granulocytes |
| Zdroj: | Molecular and Cellular Probes. 12:93-99 |
| ISSN: | 0890-8508 |
| DOI: | 10.1006/mcpr.1998.0150 |
| Popis: | The potential of fatal outcome for patients afflicted with human ehrlichioses (HME and HGE) necessitates fast and accurate detection of the aetiologic agents and timely antibiotic treatment. A polymerase chain reaction (PCR)-based protocol is described that can detect as little as 10 copies of ehrlichial 16S rDNA and as few as 0·3 HGE-infected neutrophils. The method employs DNAzol for rapid DNA extraction from unfractionated whole blood in less than 1 h. For DNA amplification, highly specific oligonucleotide primers are designed that efficiently detect and distinguish between Ehrlichia chaffeensis and the HGE agent. These primers do not prime DNA extracted from closely related ehrlichial and rickettsial species. Although total DNA extracted from human blood contains nucleic acids that can be non-specifically amplified at moderate to high MgCl 2 concentrations, such non-specific priming of non-ehrlichial DNA can be completely eliminated by lowering the MgCl 2 concentration to 1 m m . Thus, this PCR-based procedure can detect and differentiate HGE and HME with speed, simplicity, specificity and sensitivity. |
| Databáze: | OpenAIRE |
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