Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene

C. Kuyler Doyle, Marcelo B. Labruna, Edward B. Breitschwerdt, Yi Wei Tang, Richard E. Corstvet, Barbara C. Hegarty, Karen C. Bloch, Ping Li, David Walker, Jere McBride

Research output: Contribution to journalArticle

116 Citations (Scopus)

Abstract

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 108 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.

Original languageEnglish (US)
Pages (from-to)504-510
Number of pages7
JournalJournal of Molecular Diagnostics
Volume7
Issue number4
StatePublished - Oct 2005

Fingerprint

Ehrlichia
Real-Time Polymerase Chain Reaction
Ehrlichia chaffeensis
Ehrlichia canis
Genes
Rickettsia typhi
Rickettsia conorii
Anaplasma
Anaplasma phagocytophilum
Ehrlichiosis
Species Specificity
DNA Primers
Arthropods
Pets
Zoonoses
Livestock
Ticks
Infection
Disulfides
Canidae

ASJC Scopus subject areas

  • Molecular Biology

Cite this

Doyle, C. K., Labruna, M. B., Breitschwerdt, E. B., Tang, Y. W., Corstvet, R. E., Hegarty, B. C., ... McBride, J. (2005). Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene. Journal of Molecular Diagnostics, 7(4), 504-510.

Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene. / Doyle, C. Kuyler; Labruna, Marcelo B.; Breitschwerdt, Edward B.; Tang, Yi Wei; Corstvet, Richard E.; Hegarty, Barbara C.; Bloch, Karen C.; Li, Ping; Walker, David; McBride, Jere.

In: Journal of Molecular Diagnostics, Vol. 7, No. 4, 10.2005, p. 504-510.

Research output: Contribution to journalArticle

Doyle, CK, Labruna, MB, Breitschwerdt, EB, Tang, YW, Corstvet, RE, Hegarty, BC, Bloch, KC, Li, P, Walker, D & McBride, J 2005, 'Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene', Journal of Molecular Diagnostics, vol. 7, no. 4, pp. 504-510.
Doyle, C. Kuyler ; Labruna, Marcelo B. ; Breitschwerdt, Edward B. ; Tang, Yi Wei ; Corstvet, Richard E. ; Hegarty, Barbara C. ; Bloch, Karen C. ; Li, Ping ; Walker, David ; McBride, Jere. / Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene. In: Journal of Molecular Diagnostics. 2005 ; Vol. 7, No. 4. pp. 504-510.
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