Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene
| Autor: | Jere W. McBride, Karen C. Bloch, C. Kuyler Doyle, Richard E. Corstvet, Barbara C. Hegarty, Edward B. Breitschwerdt, Ping Li, Marcelo Bahia Labruna, David H. Walker, Yi-Wei Tang |
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| Rok vydání: | 2005 |
| Předmět: |
Anaplasma platys
Ehrlichia ewingii Time Factors Ehrlichia canis animal diseases Ehrlichia Color Polymerase Chain Reaction Sensitivity and Specificity Pathology and Forensic Medicine law.invention Dogs law Rickettsia typhi RNA Ribosomal 16S parasitic diseases TaqMan Ehrlichia chaffeensis Animals Humans Taq Polymerase Polymerase chain reaction biology Ehrlichiosis Protein Disulfide Reductase (Glutathione) biology.organism_classification bacterial infections and mycoses Virology Molecular Medicine bacteria Regular Articles |
| Zdroj: | The Journal of molecular diagnostics : JMD. 7(4) |
| ISSN: | 1525-1578 |
| Popis: | Ehrlichia species are the etiological agents of emerging and life-threatening tick-borne human zoonoses, in addition to causing serious and fatal infections in companion animals and livestock. We developed the first tricolor TaqMan real-time polymerase chain reaction assay capable of simultaneously detecting and discriminating medically important ehrlichiae in a single reaction. Analytical sensitivity of 50 copies per reaction was attained with templates from Ehrlichia chaffeensis, Ehrlichia ewingii, and Ehrlichia canis by amplifying the genus-specific disulfide bond formation protein gene (dsb). Ehrlichia genus-specific dsb primers amplified DNA from all known Ehrlichia species but not from other rickettsial organisms including Anaplasma platys, Anaplasma phagocytophilum, Rickettsia conorii, or Rickettsia typhi. High species specificity was attained as each species-specific TaqMan probe (E. chaffeensis, E. ewingii, and E. canis) identified homologous templates but did not cross-hybridize with heterologous Ehrlichia templates at concentrations as high as 10(8) copies. Identification of E. chaffeensis, E. ewingii, and E. canis from natural and experimental infections, previously confirmed by polymerase chain reaction and serological or microscopic evidence, demonstrated the comparable specificity and sensitivity of the dsb real-time assay. This assay provides a powerful tool for prospective medical diagnosis for human and canine ehrlichioses and for ecologic and epidemiological studies involving arthropod and mammalian hosts. |
| Databáze: | OpenAIRE |
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