The first isolation, in vitro propagation, and genetic characterization of Ehrlichia canis in Israel

Keysary, Avi; Waner, Trevor; Rosner, Miri; Warner, Cynthia K.; Dawson, Jacqueline E.; Zass, R.; Biggie, Kristine L.; Harrus, Shimon

doi:10.1016/0304-4017(95)00866-7

Cited by 55 publications

(46 citation statements)

References 9 publications

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“…Previous studies demonstrated a higher sensitivity of Dot-ELISA when compared to IFAT, although both methods are qualitatively efficient in detecting anti-E. canis antibodies (CADMAN et al, 1994;OLIVEIRA et al, 2000;HARRUS et al, 2002). The Israeli isolate, used in Dot-ELISA, was 0.54% different from the Oklahoma strain used in IFAT (KEYSARY et al, 1996). Besides the high sensitivity, Dot-ELISA is a rapid technique and easy to be used in clinical routine for the diagnosis of ehrlichia.…”

Section: Discussionmentioning

confidence: 99%

Canine ehrlichiosis: clinical, hematological, serological and molecular aspects

et al. 2008

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Section: Discussionmentioning

confidence: 99%

Canine ehrlichiosis: clinical, hematological, serological and molecular aspects

et al. 2008

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“…Chromosomal DNA from four canine E. canis isolates was used (Florida, DJ [North Carolina; 5], Jellybean [Virginia], and Israel no. 611 [13]). Three human E. chaffeensis isolates from Florida, but from different counties (Wakulla, Liberty, and Osceola), were obtained from Roman Reddy (Kansas State University, Manhattan, Kans.).…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Major Antigenic Protein 2 of Ehrlichia canis and Ehrlichia chaffeensis and Its Application for Serodiagnosis of Ehrlichiosis

Knowles

Alleman

Sorenson

et al. 2003

Clin Vaccine Immunol

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Canine monocytic ehrlichiosis, caused by Ehrlichia canis or Ehrlichia chaffeensis, can result in clinical disease in naturally infected animals. Coinfections with these agents may be common in certain areas of endemicity. Currently, a species-specific method for serological diagnosis of monocytic ehrlichiosis is not available. Previously, we developed two indirect enzyme-linked immunosorbent assays (ELISAs) using the major antigenic protein 2 (MAP2) of E. chaffeensis and E. canis. In this study, we further characterized the conservation of MAP2 among various geographic isolates of each organism and determined if the recombinant MAP2 (rMAP2) of E. chaffeensis would cross-react with E. canis-infected dog sera. Genomic Southern blot analysis using digoxigenin-labeled species-specific probes suggested that map2 is a single-copy gene in both Ehrlichia species. Sequences of the single map2 genes of seven geographically different isolates of E. chaffeensis and five isolates of E. canis are highly conserved among the various isolates of each respective ehrlichial species. ELISA and Western blot analysis confirmed that the E. chaffeensis rMAP2 failed to serologically differentiate between E. canis and E. chaffeensis infections.

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“…7 Host cell-free E. canis suspension was prepared by homogenizing heavily infected E. canis-DH82 cells for 30 seconds on an ice bath. The cell suspension was subsequently centrifuged at 3,200 ϫ g for 5 seconds at 6 C. The host cell-free E. canis supernatant was then inoculated onto a 6-well plate confluent with J774.A1 cells and centrifuged at 3,200 ϫ g for 15 minutes at room temperature.…”

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“…7 The organisms were arranged within phagosomes in membrane-bound vacuoles (Fig: 1). The number of vacuoles and the number of organisms within the vacuoles of the J774.A1 and DH82 cells appeared to be similar.…”

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“…The ultrastructure of the erhlichial organisms was found to be similar to those described in the DH82 cell line. 7 The significance of the empty membrane packets within the phagosome is not clear but may represent a defect in the assembly of the ehrlichial organisms within the cytoplamsic vacuoles. No cytopathic effects were seen in the J774.A1 cells due to E. canis infection.…”

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Cultivation of Ehrlichia Canis in a Continuous BALB/C Mouse Macrophage Cell Culture Line

et al. 2001

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Abstract. This report describes the successful adaptation of the Israeli isolate of Ehrlichia canis on a continuous mouse macrophage cell line (J774.A1). Successful infection of the J774.A1 cells was first judged by the direct immunofluorescence antibody test using an anti-E. canis-IgG:FITC conjugate. A particular property of infected J774.A1 cells was the ability to reestablish after harvesting of the monolayer by scaping. Infected cells were used as antigen for immunofluorescence antibody tests (IFA), and the results compared well with those of DH82 cells. It was concluded that the J774.A1 continuous cell line could serve as an alternate propagation cell line for E. canis organisms.

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The first isolation, in vitro propagation, and genetic characterization of Ehrlichia canis in Israel

Cited by 55 publications

References 9 publications

Canine ehrlichiosis: clinical, hematological, serological and molecular aspects

Canine ehrlichiosis: clinical, hematological, serological and molecular aspects

Characterization of the Major Antigenic Protein 2 of Ehrlichia canis and Ehrlichia chaffeensis and Its Application for Serodiagnosis of Ehrlichiosis

Cultivation of Ehrlichia Canis in a Continuous BALB/C Mouse Macrophage Cell Culture Line

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