This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Massung, R. F.
Right arrow Articles by Olson, J. G.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Massung, R. F.
Right arrow Articles by Olson, J. G.

 Previous Article  |  Next Article 

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.


This article has been cited by other articles:

  • Zeman, P., Jahn, P. (2009). An entropy-optimized multilocus approach for characterizing the strains of Anaplasma phagocytophilum infecting horses in the Czech Republic. J Med Microbiol 58: 423-429 [Abstract] [Full Text]  
  • Santos, A. S., Alexandre, N., Sousa, R., Nuncio, M. S., Bacellar, F., Dumler, J. S. (2009). Serological and molecular survey of Anaplasma species infection in dogs with suspected tickborne disease in Portugal. Vet Rec. 164: 168-171 [Abstract] [Full Text]  
  • Ladbury, G. A. F., Stuen, S., Thomas, R., Bown, K. J., Woldehiwet, Z., Granquist, E. G., Bergstrom, K., Birtles, R. J. (2008). Dynamic Transmission of Numerous Anaplasma phagocytophilum Genotypes among Lambs in an Infected Sheep Flock in an Area of Anaplasmosis Endemicity. J. Clin. Microbiol. 46: 1686-1691 [Abstract] [Full Text]  
  • Massung, R. F., Levin, M. L., Munderloh, U. G., Silverman, D. J., Lynch, M. J., Gaywee, J. K., Kurtti, T. J. (2007). Isolation and Propagation of the Ap-Variant 1 Strain of Anaplasma phagocytophilum in a Tick Cell Line. J. Clin. Microbiol. 45: 2138-2143 [Abstract] [Full Text]  
  • Oteo, J. A., Portillo, A., Santibanez, S., Blanco, J. R., Perez-Martinez, L., Ibarra, V. (2006). Cluster of Cases of Human Rickettsia felis Infection from Southern Europe (Spain) Diagnosed by PCR.. J. Clin. Microbiol. 44: 2669-2671 [Abstract] [Full Text]  
  • Kawahara, M., Rikihisa, Y., Lin, Q., Isogai, E., Tahara, K., Itagaki, A., Hiramitsu, Y., Tajima, T. (2006). Novel Genetic Variants of Anaplasma phagocytophilum, Anaplasma bovis, Anaplasma centrale, and a Novel Ehrlichia sp. in Wild Deer and Ticks on Two Major Islands in Japan. Appl. Environ. Microbiol. 72: 1102-1109 [Abstract] [Full Text]  
  • Massung, R. F., Mather, T. N., Levin, M. L. (2006). Reservoir Competency of Goats for the Ap-Variant 1 Strain of Anaplasma phagocytophilum. Infect. Immun. 74: 1373-1375 [Abstract] [Full Text]  
  • de la Fuente, J., Massung, R. F., Wong, S. J., Chu, F. K., Lutz, H., Meli, M., von Loewenich, F. D., Grzeszczuk, A., Torina, A., Caracappa, S., Mangold, A. J., Naranjo, V., Stuen, S., Kocan, K. M. (2005). Sequence Analysis of the msp4 Gene of Anaplasma phagocytophilum Strains. J. Clin. Microbiol. 43: 1309-1317 [Abstract] [Full Text]  
  • Lin, Q., Rikihisa, Y., Massung, R. F., Woldehiwet, Z., Falco, R. C. (2004). Polymorphism and Transcription at the p44-1/p44-18 Genomic Locus in Anaplasma phagocytophilum Strains from Diverse Geographic Regions. Infect. Immun. 72: 5574-5581 [Abstract] [Full Text]  
  • Courtney, J. W., Kostelnik, L. M., Zeidner, N. S., Massung, R. F. (2004). Multiplex Real-Time PCR for Detection of Anaplasma phagocytophilum and Borrelia burgdorferi. J. Clin. Microbiol. 42: 3164-3168 [Abstract] [Full Text]  
  • Casey, A. N. J., Birtles, R. J., Radford, A. D., Bown, K. J., French, N. P., Woldehiwet, Z., Ogden, N. H. (2004). Groupings of highly similar major surface protein (p44)-encoding paralogues: a potential index of genetic diversity amongst isolates of Anaplasma phagocytophilum. Microbiology 150: 727-734 [Abstract] [Full Text]  
  • Benson, M. J., Gawronski, J. D., Eveleigh, D. E., Benson, D. R. (2004). Intracellular Symbionts and Other Bacteria Associated with Deer Ticks (Ixodes scapularis) from Nantucket and Wellfleet, Cape Cod, Massachusetts. Appl. Environ. Microbiol. 70: 616-620 [Abstract] [Full Text]  
  • Liddell, A. M., Stockham, S. L., Scott, M. A., Sumner, J. W., Paddock, C. D., Gaudreault-Keener, M., Arens, M. Q., Storch, G. A. (2003). Predominance of Ehrlichia ewingii in Missouri Dogs. J. Clin. Microbiol. 41: 4617-4622 [Abstract] [Full Text]  
  • CAO, W.-C., ZHAO, Q.-M., ZHANG, P.-H., YANG, H., WU, X.-M., WEN, B.-H., ZHANG, X.-T., HABBEMA, J. D. F. (2003). PREVALENCE OF ANAPLASMA PHAGOCYTOPHILA AND BORRELIA BURGDORFERI IN IXODES PERSULCATUS TICKS FROM NORTHEASTERN CHINA. Am J Trop Med Hyg 68: 547-550 [Abstract] [Full Text]  
  • Ogden, N. H., Casey, A. N. J., Woldehiwet, Z., French, N. P. (2003). Transmission of Anaplasma phagocytophilum to Ixodes ricinus Ticks from Sheep in the Acute and Post-Acute Phases of Infection. Infect. Immun. 71: 2071-2078 [Abstract] [Full Text]  
  • Courtney, J. W., Dryden, R. L., Montgomery, J., Schneider, B. S., Smith, G., Massung, R. F. (2003). Molecular Characterization of Anaplasma phagocytophilum and Borrelia burgdorferi in Ixodes scapularis Ticks from Pennsylvania. J. Clin. Microbiol. 41: 1569-1573 [Abstract] [Full Text]  
  • Arens, M. Q., Liddell, A. M., Buening, G., Gaudreault-Keener, M., Sumner, J. W., Comer, J. A., Buller, R. S., Storch, G. A. (2003). Detection of Ehrlichia spp. in the Blood of Wild White-Tailed Deer in Missouri by PCR Assay and Serologic Analysis. J. Clin. Microbiol. 41: 1263-1265 [Abstract] [Full Text]  
  • Massung, R. F., Slater, K. G. (2003). Comparison of PCR Assays for Detection of the Agent of Human Granulocytic Ehrlichiosis, Anaplasma phagocytophilum. J. Clin. Microbiol. 41: 717-722 [Abstract] [Full Text]  
  • Stuen, S., Van De Pol, I., Bergstrom, K., Schouls, L. M. (2002). Identification of Anaplasma phagocytophila (Formerly Ehrlichia phagocytophila) Variants in Blood from Sheep in Norway. J. Clin. Microbiol. 40: 3192-3197 [Abstract] [Full Text]  
  • Bjoersdorff, A., Bagert, B., Massung, R. F., Gusa, A., Eliasson, I. (2002). Isolation and Characterization of Two European Strains of Ehrlichia phagocytophila of Equine Origin. CVI 9: 341-343 [Abstract] [Full Text]  
  • Liz, J. S., Sumner, J. W., Pfister, K., Brossard, M. (2002). PCR Detection and Serological Evidence of Granulocytic Ehrlichial Infection in Roe Deer (Capreolus capreolus) and Chamois (Rupicapra rupicapra). J. Clin. Microbiol. 40: 892-897 [Abstract] [Full Text]  
  • Gusa, A. A., Buller, R. S., Storch, G. A., Huycke, M. M., Machado, L. J., Slater, L. N., Stockham, S. L., Massung, R. F. (2001). Identification of a p28 Gene in Ehrlichia ewingii: Evaluation of Gene for Use as a Target for a Species-Specific PCR Diagnostic Assay. J. Clin. Microbiol. 39: 3871-3876 [Abstract] [Full Text]  
  • Cao, W.-C., Zhao, Q.-M., Zhang, P.-H., Dumler, J. S., Zhang, X.-T., Fang, L.-Q., Yang, H. (2000). Granulocytic Ehrlichiae in Ixodes persulcatus Ticks from an Area in China Where Lyme Disease Is Endemic. J. Clin. Microbiol. 38: 4208-4210 [Abstract] [Full Text]  
  • Wolf, L., McPherson, T., Harrison, B., Engber, B., Anderson, A., Whitt, P. (2000). Prevalence of Ehrlichia ewingii in Amblyomma americanum in North Carolina. J. Clin. Microbiol. 38: 2795-2795 [Full Text]  
  • Liz, J. S., Anderes, L., Sumner, J. W., Massung, R. F., Gern, L., Rutti, B., Brossard, M. (2000). PCR Detection of Granulocytic Ehrlichiae in Ixodes ricinus Ticks and Wild Small Mammals in Western Switzerland. J. Clin. Microbiol. 38: 1002-1007 [Abstract] [Full Text]  
  • Baumgarten, B. U., Röllinghoff, M., Bogdan, C. (1999). Prevalence of Borrelia burgdorferi and Granulocytic and Monocytic Ehrlichiae in Ixodes ricinus Ticks from Southern Germany. J. Clin. Microbiol. 37: 3448-3451 [Abstract] [Full Text]  
  • Stafford, K. C. III, Massung, R. F., Magnarelli, L. A., Ijdo, J. W., Anderson, J. F. (1999). Infection with Agents of Human Granulocytic Ehrlichiosis, Lyme Disease, and Babesiosis in Wild White-Footed Mice (Peromyscus leucopus) in Connecticut. J. Clin. Microbiol. 37: 2887-2892 [Abstract] [Full Text]  
  • Childs, J. E., Sumner, J. W., Nicholson, W. L., Massung, R. F., Standaert, S. M., Paddock, C. D. (1999). Outcome of Diagnostic Tests Using Samples from Patients with Culture-Proven Human Monocytic Ehrlichiosis: Implications for Surveillance. J. Clin. Microbiol. 37: 2997-3000 [Abstract] [Full Text]  
  • Schouls, L. M., Van De Pol, I., Rijpkema, S. G. T., Schot, C. S. (1999). Detection and Identification of Ehrlichia, Borrelia burgdorferi Sensu Lato, and Bartonella Species in Dutch Ixodes ricinus Ticks. J. Clin. Microbiol. 37: 2215-2222 [Abstract] [Full Text]  
  • Comer, J. A., Nicholson, W. L., Sumner, J. W., Olson, J. G., Childs, J. E. (1999). Diagnosis of Human Ehrlichiosis by PCR Assay of Acute-Phase Serum. J. Clin. Microbiol. 37: 31-34 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»