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Primers were used to amplify a 561-bp region of the 16S rRNA gene of Ehrlichia phagocytophila fro... more Primers were used to amplify a 561-bp region of the 16S rRNA gene of Ehrlichia phagocytophila from Ixodes scapularis ticks and small mammals collected in Rhode Island and Connecticut. DNA sequences for all 50 E. phagocytophila-positive samples collected from 1996 through 1998 in southwestern Connecticut were identical to the sequence reported for E. phagocytophila DNA from confirmed human cases. In contrast, the sequences from 92 of 123 E. phagocytophila-positive Rhode Island samples collected from 1996 through 1999 included several variants differing by 1-2 nucleotides from that in the agent infecting humans. While 11.9% of 67 E. phagocytophila-positive ticks collected during 1997 in Rhode Island harbored ehrlichiae with sequences identical to that of the human agent, 79.1% had a variant sequence not previously described. The low incidence of human ehrlichiosis in Rhode Island may in part result from these variant ehrlichiae's interference with the maintenance and transmission of the true agent of human disease.
A sensitive and specific nested PCR assay was developed for the detection of granulocytic ehrlich... more A sensitive and specific nested PCR assay was developed for the detection of granulocytic ehrlichiae. The assay amplifies the 16S rRNA gene and was used to examine acute-phase EDTA-blood and serum samples obtained from seven humans with clinical presentations compatible with human granulocytic ehrlichiosis. Five of the seven suspected cases were positive by the PCR assay using DNA extracted from whole blood as the template, compared with a serologic assay that identified only one positive sample. The PCR assay using DNA extracted from the corresponding serum samples as the template identified three positive samples. The sensitivity of the assay on human samples was examined, and the limit of detection was shown to be fewer than 2 copies of the 16S rRNA gene. The application of the assay to nonhuman samples demonstrated products amplified from template DNA extracted from Ixodes scapularis ticks collected in Rhode Island and from EDTA-blood specimens obtained from white-tailed deer in Maryland. All PCR products were sequenced and identified as specific to granulocytic ehrlichiae. A putative variant granulocytic ehrlichia 16S rRNA gene sequence was detected among products amplified from both the ticks and the deer blood specimens.
Journal of Small Animal Practice, 1999
A 14-month-old shorthaired cat was presented to the Animal Hospital in Skara, Sweden, with a two-... more A 14-month-old shorthaired cat was presented to the Animal Hospital in Skara, Sweden, with a two-day history of lethargy, anorexia and tachypnoea. Clinical examination and laboratory investigations revealed fever, dehydration, tick infestation, neutrophilia with left shift, lymphopenia, hyperglycaemia and intracytoplasmic neutrophilic Ehrlichia inclusions. After treatment with intravenous doxycycline and lactated Ringer's solution the temperature returned to normal. Oral treatment with doxycycline continued for 20 days. The ehrlichiosis diagnosis was confirmed by serology, polymerase chain reaction and DNA sequencing. No relapse was observed during the eight-month follow-up period. The granulocytotropic Ehrlichia strain found in the cat was later characterised by analysis of the 16S rRNA gene sequence which showed 100 per cent identity to DNA sequences found in Swedish canine and equine granulocytotropic Ehrlichia strains. This is, to the best of the authors' knowledge, the first reported case of granulocytic ehrlichiosis in a cat.
Primers were used to amplify a 561-bp region of the 16S rRNA gene of Ehrlichia phagocytophila fro... more Primers were used to amplify a 561-bp region of the 16S rRNA gene of Ehrlichia phagocytophila from Ixodes scapularis ticks and small mammals collected in Rhode Island and Connecticut. DNA sequences for all 50 E. phagocytophila-positive samples collected from 1996 through 1998 in southwestern Connecticut were identical to the sequence reported for E. phagocytophila DNA from confirmed human cases. In contrast, the sequences from 92 of 123 E. phagocytophila-positive Rhode Island samples collected from 1996 through 1999 included several variants differing by 1-2 nucleotides from that in the agent infecting humans. While 11.9% of 67 E. phagocytophila-positive ticks collected during 1997 in Rhode Island harbored ehrlichiae with sequences identical to that of the human agent, 79.1% had a variant sequence not previously described. The low incidence of human ehrlichiosis in Rhode Island may in part result from these variant ehrlichiae's interference with the maintenance and transmission of the true agent of human disease.
A sensitive and specific nested PCR assay was developed for the detection of granulocytic ehrlich... more A sensitive and specific nested PCR assay was developed for the detection of granulocytic ehrlichiae. The assay amplifies the 16S rRNA gene and was used to examine acute-phase EDTA-blood and serum samples obtained from seven humans with clinical presentations compatible with human granulocytic ehrlichiosis. Five of the seven suspected cases were positive by the PCR assay using DNA extracted from whole blood as the template, compared with a serologic assay that identified only one positive sample. The PCR assay using DNA extracted from the corresponding serum samples as the template identified three positive samples. The sensitivity of the assay on human samples was examined, and the limit of detection was shown to be fewer than 2 copies of the 16S rRNA gene. The application of the assay to nonhuman samples demonstrated products amplified from template DNA extracted from Ixodes scapularis ticks collected in Rhode Island and from EDTA-blood specimens obtained from white-tailed deer in Maryland. All PCR products were sequenced and identified as specific to granulocytic ehrlichiae. A putative variant granulocytic ehrlichia 16S rRNA gene sequence was detected among products amplified from both the ticks and the deer blood specimens.
Journal of Small Animal Practice, 1999
A 14-month-old shorthaired cat was presented to the Animal Hospital in Skara, Sweden, with a two-... more A 14-month-old shorthaired cat was presented to the Animal Hospital in Skara, Sweden, with a two-day history of lethargy, anorexia and tachypnoea. Clinical examination and laboratory investigations revealed fever, dehydration, tick infestation, neutrophilia with left shift, lymphopenia, hyperglycaemia and intracytoplasmic neutrophilic Ehrlichia inclusions. After treatment with intravenous doxycycline and lactated Ringer's solution the temperature returned to normal. Oral treatment with doxycycline continued for 20 days. The ehrlichiosis diagnosis was confirmed by serology, polymerase chain reaction and DNA sequencing. No relapse was observed during the eight-month follow-up period. The granulocytotropic Ehrlichia strain found in the cat was later characterised by analysis of the 16S rRNA gene sequence which showed 100 per cent identity to DNA sequences found in Swedish canine and equine granulocytotropic Ehrlichia strains. This is, to the best of the authors' knowledge, the first reported case of granulocytic ehrlichiosis in a cat.