title: Isolation and characterization of Ehrlichia chaffeensis strains from patients with fatal ehrlichiosis.
abstract: Two new isolates of Ehrlichia chaffeensis (designated Jax and St. Vincent) were obtained from patients with fatal ehrlichial infections. Patients developed characteristic manifestations of severe disease due to E. chaffeensis, including marked thrombocytopenia, pulmonary insufficiency, and encephalopathy. Primary isolation was achieved in DH82 cells; the Jax and St. Vincent isolates were detected within 19 and 8 days postinoculation, respectively. The isolates were characterized by molecular evaluation of the 16S rRNA gene, the groESL heat shock operon, a 120-kDa immunodominant protein gene, and an incompletely characterized repetitive-motif sequence (variable-length PCR target [VLPT]). The sequences were compared with those of the corresponding molecular regions in the type isolate (Arkansas). St. Vincent contained one fewer repeat unit in both the 120-kDa protein gene and the VLPT compared with corresponding sequences of the Jax and Arkansas isolates. 16S rRNA gene sequences from the two new isolates had 100% identity to the corresponding sequences of the 91HE17 and Sapulpa isolates of E. chaffeensis, and to the corrected 16S rRNA gene sequence of the Arkansas isolate. The Jax isolate grew more slowly than the St. Vincent isolate in DH82 cells, and both of the new isolates grew more slowly than the extensively passaged Arkansas isolate. Although specific associations between ehrlichial pathogenicity and genotype were not identified from these comparisons, recovery of this organism from a spectrum of clinical presentations remains an integral step in understanding mechanisms of disease caused by E. chaffeensis.
authors.affiliation: Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA. cdp9@cdc.gov
authors.forename: C D
authors.initials: CD
authors.lastname: Paddock
authors.forename: J W
authors.initials: JW
authors.lastname: Sumner
authors.forename: G M
authors.initials: GM
authors.lastname: Shore
authors.forename: D C
authors.initials: DC
authors.lastname: Bartley
authors.forename: R C
authors.initials: RC
authors.lastname: Elie
authors.forename: J G
authors.initials: JG
authors.lastname: McQuade
authors.forename: C R
authors.initials: CR
authors.lastname: Martin
authors.forename: C S
authors.initials: CS
authors.lastname: Goldsmith
authors.forename: J E
authors.initials: JE
authors.lastname: Childs
mesh_terms: D000704:Analysis of Variance; D000942:Antigens, Bacterial / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics; D018835:Chaperonin 10 / Q000235:genetics; D018834:Chaperonin 60 / Q000235:genetics; D018469:Ehrlichia chaffeensis / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D016873:Ehrlichiosis / Q000097:blood / Q000209:etiology / Q000382:microbiology* / Q000401:mortality; D005260:Female; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D017422:Sequence Analysis, DNA
chemical_list: C104346:120-kDa immunodominant protein, Ehrlichia chaffeensis; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; D016056:Immunodominant Epitopes; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S