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Journal of Clinical Microbiology, April 1998, p. 1090-1095, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Nested PCR Assay for Detection of
Granulocytic Ehrlichiae
Robert F.
Massung,1,*
Kim
Slater,1
Jessica H.
Owens,1
William L.
Nicholson,1
Thomas N.
Mather,2
Victoria B.
Solberg,3 and
James G.
Olson1
National Center for Infectious Diseases,
Centers for Disease Control and Prevention, Atlanta, Georgia
303331;
Center for Vector-Borne
Disease, University of Rhode Island, Kingston, Rhode Island
028812; and
Department of
Entomology, Walter Reed Army Institute of Research, Washington,
D.C. 20307-51003
Received 6 August 1997/Returned for modification 10 November
1997/Accepted 12 January 1998
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.
*
Corresponding author. Mailing address: Centers for
Disease Control and Prevention, 1600 Clifton Rd., MS G-13, Atlanta, GA 30333. Phone: (404) 639-1075. Fax: (404) 639-4436. E-mail:
rfm2{at}cdc.gov.
Journal of Clinical Microbiology, April 1998, p. 1090-1095, Vol. 36, No. 4
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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