Affiliation:
1. School of Medicine, University of Maryland—Baltimore, Baltimore, Maryland 21201-1559
2. Viral and Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333
Abstract
ABSTRACT
A spotted fever rickettsia quantitative PCR assay (SQ-PCR) was developed for the detection and enumeration of
Rickettsia rickettsii
and other closely related spotted fever group rickettsiae. The assay is based on fluorescence detection of SYBR Green dye intercalation in a 154-bp fragment of the rOmpA gene during amplification by PCR. As few as 5 copies of the rOmpA gene of
R. rickettsii
can be detected. SQ-PCR is suitable for quantitation of
R. rickettsii
and 10 other genotypes of spotted fever group rickettsiae but not for
R. akari
,
R. australis
,
R. bellii
, or typhus group rickettsiae. The sensitivity of SQ-PCR was comparable to that of a plaque assay using centrifugation for inoculation. The SQ-PCR assay was applied successfully to the characterization of rickettsial stock cultures, the replication of rickettsiae in cell culture, the recovery of rickettsial DNA following different methods of extraction, and the quantitation of rickettsial loads in infected animal tissues, clinical samples, and ticks.
Publisher
American Society for Microbiology
Reference24 articles.
1. A protective protein antigen of Rickettsia rickettsii has tandemly repeated, near-identical sequences
2. Dasch G. A. and E. Weiss. 1992. The genera Rickettsia Rochalimaea Ehrlichia Cowdria and Neorickettsia p. 2407-2470. In A. Balows H. G. Truper M. Dworkin W. Harder and K. H. Schleifer (ed.) The prokaryotes. A handbook on the biology of bacteria: ecophysiology isolation identification applications 2nd ed. vol. III. Springer-Verlag New York N.Y.
3. Emelyanov, V. V. 1990. A simple method for counting Rickettsia cells. Mikrobiol. Z.52:91-95.
4. Eremeeva, M. E., Z. Liang, C. Paddock, S. Zaki, J. G. Vandenbergh, G. A. Dasch, and D. J. Silverman. 2003. Rickettsia rickettsii infection in the pine vole, Microtus pinetorum: kinetics of infection and quantitation of antioxidant enzyme gene expression by RT-PCR. Ann. N. Y. Acad. Sci.990:468-473.
5. Eremeeva M. E. and G. A. Dasch. 2001. Rickettsia and Orientia p. 2175-2216. In M. Sussman (ed.) Molecular medical microbiology vol. 3. Academic Press London United Kingdom.
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