Skip to main content
SpringerLink
Log in

Rickettsiales: Laboratory Diagnosis

  • Chapter
  • First Online:
Rickettsiales

  • 768 Accesses

Abstract

As undifferentiated febrile illnesses, infections caused by organisms in the genus Rickettsia, Orientia, Ehrlichia, and Anaplasma are often difficult to diagnose. Clinical suspicion is the key to the initiation of timely therapy and clinical workup. Serology is the mainstay of diagnostic methods, but since antibody production occurs later in illness, diagnosis is usually retrospective. Serologic confirmation requires seroconversion or a fourfold increase in titers from acute- and convalescent-phase sera. Immunohistochemistry offers a method of diagnosis during acute illness, but this technique is not readily available to most physicians. Molecular methods also offer the chance of making a diagnosis during early illness, but the sensitivity of these assays is dependent on the pathogenesis of the infecting organism and the site of DNA acquisition (e.g., blood, buffy coat, tissue).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  • Aguero-Rosenfeld ME, Donnarumma L, Zentmaier L, Jacob J, Frey M, et al. (2002) Seroprevalence of antibodies that react with Anaplasma phagocytophila, the agent of human granulocytic ehrlichiosis, in different populations in Westchester County, New York. J Clin Microbiol 40: 2612-2615.

    Article  PubMed  PubMed Central  Google Scholar 

  • Allerdice ME, Pritt BS, Sloan LM, Paddock CD, Karpathy SE (2016) A real-time PCR assay for detection of the Ehrlichia muris-like agent, a newly recognized pathogen of humans in the upper Midwestern United States. Ticks Tick Borne Dis 7: 146-149.

    Article  PubMed  Google Scholar 

  • Bakken JS, Aguero-Rosenfeld ME, Tilden RL, Wormser GP, Horowitz HW, et al. (2001) Serial measurements of hematologic counts during the active phase of human granulocytic ehrlichiosis. Clin Infect Dis 32: 862-870.

    Article  CAS  PubMed  Google Scholar 

  • Bakken JS, Haller I, Riddell D, Walls JJ, Dumler JS (2002) The serological response of patients infected with the agent of human granulocytic ehrlichiosis. Clin Infect Dis 34: 22-27.

    Article  PubMed  Google Scholar 

  • Bakken JS, Krueth J, Wilson-Nordskog C, Tilden RL, Asanovich K, et al. (1996) Clinical and laboratory characteristics of human granulocytic ehrlichiosis. JAMA 275: 199-205.

    Article  CAS  PubMed  Google Scholar 

  • Blacksell SD, Bryant NJ, Paris DH, Doust JA, Sakoda Y, et al. (2007) Scrub typhus serologic testing with the indirect immunofluorescence method as a diagnostic gold standard: a lack of consensus leads to a lot of confusion. Clin Infect Dis 44: 391-401.

    Article  PubMed  Google Scholar 

  • Blanton LS (2013) Rickettsial infections in the tropics and in the traveler. Curr Opin Infect Dis 26: 435-440.

    Article  PubMed  Google Scholar 

  • Blanton LS, Lea AS, Kelly BC, Walker DH (2015) An unusual cutaneous manifestation in a patient with murine typhus. Am J Trop Med Hyg 93: 1164-1167.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Blanton LS, Walker DH, Bouyer DH (2014) Rickettsiae and ehrlichiae within a city park: is the urban dweller at risk? Vector Borne Zoonotic Dis 14: 168-170.

    Article  PubMed  Google Scholar 

  • Boostrom A, Beier MS, Macaluso JA, Macaluso KR, Sprenger D, et al. (2002) Geographic association of Rickettsia felis-infected opossums with human murine typhus, Texas. Emerg Infect Dis 8: 549-554.

    Article  PubMed  PubMed Central  Google Scholar 

  • Bozeman FM, Elisberg BL (1963) Serological diagnosis of scrub typhus by indirect immunofluorescence. Proc Soc Exp Biol Med 112: 568-573.

    Article  CAS  PubMed  Google Scholar 

  • Chapman AS, Bakken JS, Folk SM, Paddock CD, Bloch KC, et al. (2006) Diagnosis and management of tickborne rickettsial diseases: Rocky Mountain spotted fever, ehrlichioses, and anaplasmosis--United States: a practical guide for physicians and other health-care and public health professionals. MMWR Recomm Rep 55: 1-27.

    Google Scholar 

  • Childs JE, Sumner JW, Nicholson WL, Massung RF, Standaert SM, et al. (1999) Outcome of diagnostic tests using samples from patients with culture-proven human monocytic ehrlichiosis: implications for surveillance. J Clin Microbiol 37: 2997-3000.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Civen R, Ngo V (2008) Murine typhus: an unrecognized suburban vectorborne disease. Clin Infect Dis 46: 913-918.

    Article  PubMed  Google Scholar 

  • Clements ML, Dumler JS, Fiset P, Wisseman CL, Jr., Snyder MJ, et al. (1983) Serodiagnosis of Rocky Mountain spotted fever: comparison of IgM and IgG enzyme-linked immunosorbent assays and indirect fluorescent antibody test. J Infect Dis 148: 876-880.

    Article  CAS  PubMed  Google Scholar 

  • Dawson J, Fishbein D, Eng T, Redus M, Greene N (1990) Diagnosis of human ehrlichiosis with the indirect fluorescent antibody test: kinetics and specificity. Ann N Y Acad Sci 590: 308.

    Article  CAS  PubMed  Google Scholar 

  • Dawson JE, Paddock CD, Warner CK, Greer PW, Bartlett JH, et al. (2001) Tissue diagnosis of Ehrlichia chaffeensis in patients with fatal ehrlichiosis by use of immunohistochemistry, in situ hybridization, and polymerase chain reaction. Am J Trop Med Hyg 65: 603-609.

    CAS  PubMed  Google Scholar 

  • Doyle CK, Labruna MB, Breitschwerdt EB, Tang YW, Corstvet RE, et al. (2005) Detection of medically important Ehrlichia by quantitative multicolor TaqMan real-time polymerase chain reaction of the dsb gene. J Mol Diagn 7: 504-510.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Dumler JS, Dawson JE, Walker DH (1993) Human ehrlichiosis: hematopathology and immunohistologic detection of Ehrlichia chaffeensis. Hum Pathol 24: 391-396.

    Article  CAS  PubMed  Google Scholar 

  • Dumler JS, Taylor JP, Walker DH (1991) Clinical and laboratory features of murine typhus in south Texas, 1980 through 1987. JAMA 266: 1365-1370.

    Article  CAS  PubMed  Google Scholar 

  • Dunn BE, Monson TP, Dumler JS, Morris CC, Westbrook AB, et al. (1992) Identification of Ehrlichia chaffeensis morulae in cerebrospinal fluid mononuclear cells. J Clin Microbiol 30: 2207-2210.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Edelman DC, Dumler JS (1996) Evaluation of an Improved PCR Diagnostic Assay for Human Granulocytic Ehrlichiosis. Mol Diagn 1: 41-49.

    Article  PubMed  Google Scholar 

  • Eremeeva M, Yu X, Raoult D (1994) Differentiation among spotted fever group rickettsiae species by analysis of restriction fragment length polymorphism of PCR-amplified DNA. J Clin Microbiol 32: 803-810.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Eremeeva ME, Balayeva NM, Raoult D (1994) Serological response of patients suffering from primary and recrudescent typhus: comparison of complement fixation reaction, Weil-Felix test, microimmunofluorescence, and immunoblotting. Clin Diagn Lab Immunol 1: 318-324.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Fournier PE, Roux V, Raoult D (1998) Phylogenetic analysis of spotted fever group rickettsiae by study of the outer surface protein rOmpA. Int J Syst Bacteriol 48 Pt 3: 839-849.

    Article  CAS  PubMed  Google Scholar 

  • Horowitz HW, Aguero-Rosenfeld ME, McKenna DF, Holmgren D, Hsieh TC, et al. (1998) Clinical and laboratory spectrum of culture-proven human granulocytic ehrlichiosis: comparison with culture-negative cases. Clin Infect Dis 27: 1314-1317.

    Article  CAS  PubMed  Google Scholar 

  • Jensenius M, Fournier PE, Kelly P, Myrvang B, Raoult D (2003) African tick bite fever. Lancet Infect Dis 3: 557-564.

    Article  PubMed  Google Scholar 

  • Jensenius M, Fournier PE, Raoult D (2004) Rickettsioses and the international traveler. Clin Infect Dis 39: 1493-1499.

    Article  PubMed  Google Scholar 

  • Jiang J, Chan TC, Temenak JJ, Dasch GA, Ching WM, et al. (2004) Development of a quantitative real-time polymerase chain reaction assay specific for Orientia tsutsugamushi. Am J Trop Med Hyg 70: 351-356.

    CAS  PubMed  Google Scholar 

  • Kaplan JE, Schonberger LB (1986) The sensitivity of various serologic tests in the diagnosis of Rocky Mountain spotted fever. Am J Trop Med Hyg 35: 840-844.

    CAS  PubMed  Google Scholar 

  • Kato CY, Chung IH, Robinson LK, Austin AL, Dasch GA, et al. (2013) Assessment of real-time PCR assay for detection of Rickettsia spp. and Rickettsia rickettsii in banked clinical samples. J Clin Microbiol 51: 314-317.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kelly DJ, Wong PW, Gan E, Lewis GE, Jr. (1988) Comparative evaluation of the indirect immunoperoxidase test for the serodiagnosis of rickettsial disease. Am J Trop Med Hyg 38:400-406.

    CAS  PubMed  Google Scholar 

  • Kim CM, Cho MK, Kim DM, Yun NR, Kim SW, et al. (2016) Accuracy of conventional PCR targeting the 16S rRNA gene with the Ot-16sRF1 and Ot-16sRR1 primers for diagnosis of scrub typhus: a case-control study. J Clin Microbiol 54: 178-179.

    Article  CAS  PubMed  Google Scholar 

  • Kim DM, Kim HL, Park CY, Yang TY, Lee JH, et al. (2006) Clinical usefulness of eschar polymerase chain reaction for the diagnosis of scrub typhus: a prospective study. Clin Infect Dis 43: 1296-1300.

    Article  PubMed  Google Scholar 

  • La Scola B, Raoult D (1996) Diagnosis of Mediterranean spotted fever by cultivation of Rickettsia conorii from blood and skin samples using the centrifugation-shell vial technique and by detection of R. conorii in circulating endothelial cells: a 6-year follow-up. J Clin Microbiol 34: 2722-2727.

    PubMed  PubMed Central  Google Scholar 

  • La Scola B, Raoult D (1997) Laboratory diagnosis of rickettsioses: current approaches to diagnosis of old and new rickettsial diseases. J Clin Microbiol 35: 2715-2727.

    PubMed  PubMed Central  Google Scholar 

  • Loftis AD, Massung RF, Levin ML (2003) Quantitative real-time PCR assay for detection of Ehrlichia chaffeensis. J Clin Microbiol 41: 3870-3872.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Marrero M, Raoult D (1989) Centrifugation-shell vial technique for rapid detection of Mediterranean spotted fever rickettsia in blood culture. Am J Trop Med Hyg 40: 197-199.

    CAS  PubMed  Google Scholar 

  • Massung RF, Slater K, Owens JH, Nicholson WL, Mather TN, et al. (1998) Nested PCR assay for detection of granulocytic ehrlichiae. J Clin Microbiol 36: 1090-1095.

    CAS  PubMed  PubMed Central  Google Scholar 

  • McQuiston JH, Wiedeman C, Singleton J, Carpenter LR, McElroy K, et al. (2014) Inadequacy of IgM antibody tests for diagnosis of Rocky Mountain Spotted Fever. Am J Trop Med Hyg 91: 767-770.

    Article  PubMed  PubMed Central  Google Scholar 

  • Mouffok N, Socolovschi C, Benabdellah A, Renvoise A, Parola P, et al. (2011) Diagnosis of rickettsioses from eschar swab samples, Algeria. Emerg Infect Dis 17: 1968-1969.

    Article  PubMed  PubMed Central  Google Scholar 

  • Paddock CD, Childs JE (2003) Ehrlichia chaffeensis: a prototypical emerging pathogen. Clin Microbiol Rev 16: 37-64.

    Article  PubMed  PubMed Central  Google Scholar 

  • Paddock CD, Sumner JW, Comer JA, Zaki SR, Goldsmith CS, et al. (2004) Rickettsia parkeri: a newly recognized cause of spotted fever rickettsiosis in the United States. Clin Infect Dis 38: 805-811.

    Article  PubMed  Google Scholar 

  • Paddock CD, Sumner JW, Shore GM, Bartley DC, Elie RC, et al. (1997) Isolation and characterization of Ehrlichia chaffeensis strains from patients with fatal ehrlichiosis. J Clin Microbiol 35: 2496-2502.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Paris DH, Aukkanit N, Jenjaroen K, Blacksell SD, Day NP (2009) A highly sensitive quantitative real-time PCR assay based on the groEL gene of contemporary Thai strains of Orientia tsutsugamushi. Clin Microbiol Infect 15: 488-495.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Paris DH, Shelite TR, Day NP, Walker DH (2013) Unresolved problems related to scrub typhus: a seriously neglected life-threatening disease. Am J Trop Med Hyg 89: 301-307.

    Article  PubMed  PubMed Central  Google Scholar 

  • Phetsouvanh R, Thojaikong T, Phoumin P, Sibounheuang B, Phommasone K, et al. (2013) Inter- and intra-operator variability in the reading of indirect immunofluorescence assays for the serological diagnosis of scrub typhus and murine typhus. Am J Trop Med Hyg 88: 932-936.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pritt BS, Sloan LM, Johnson DK, Munderloh UG, Paskewitz SM, et al. (2011) Emergence of a new pathogenic Ehrlichia species, Wisconsin and Minnesota, 2009. N Engl J Med 365: 422-429.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Raoult D, Dasch GA (1995) Immunoblot cross-reactions among Rickettsia, Proteus spp. and Legionella spp. in patients with Mediterranean spotted fever. FEMS Immunol Med Microbiol 11: 13-18.

    Article  CAS  PubMed  Google Scholar 

  • Regnery RL, Spruill CL, Plikaytis BD (1991) Genotypic identification of rickettsiae and estimation of intraspecies sequence divergence for portions of two rickettsial genes. J Bacteriol 173: 1576-1589.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Roux V, Fournier PE, Raoult D (1996) Differentiation of spotted fever group rickettsiae by sequencing and analysis of restriction fragment length polymorphism of PCR-amplified DNA of the gene encoding the protein rOmpA. J Clin Microbiol 34: 2058-2065.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Roux V, Raoult D (2000) Phylogenetic analysis of members of the genus Rickettsia using the gene encoding the outer-membrane protein rOmpB (ompB). Int J Syst Evol Microbiol 50 Pt 4: 1449-1455.

    Article  CAS  PubMed  Google Scholar 

  • Salgo MP, Telzak EE, Currie B, Perlman DC, Litman N, et al. (1988) A focus of Rocky Mountain spotted fever within New York City. N Engl J Med 318: 1345-1348.

    Article  CAS  PubMed  Google Scholar 

  • Schotthoefer AM, Meece JK, Ivacic LC, Bertz PD, Zhang K, et al. (2013) Comparison of a real-time PCR method with serology and blood smear analysis for diagnosis of human anaplasmosis: importance of infection time course for optimal test utilization. J Clin Microbiol 51: 2147-2153.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sexton DJ, Kanj SS, Wilson K, Corey GR, Hegarty BC, et al. (1994) The use of a polymerase chain reaction as a diagnostic test for Rocky Mountain spotted fever. Am J Trop Med Hyg 50: 59-63.

    CAS  PubMed  Google Scholar 

  • Standaert SM, Yu T, Scott MA, Childs JE, Paddock CD, et al. (2000) Primary isolation of Ehrlichia chaffeensis from patients with febrile illnesses: clinical and molecular characteristics. J Infect Dis 181: 1082-1088.

    Article  CAS  PubMed  Google Scholar 

  • Tzianabos T, Anderson BE, McDade JE (1989) Detection of Rickettsia rickettsii DNA in clinical specimens by using polymerase chain reaction technology. J Clin Microbiol 27:2866-2868.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Walker DH (1995) Rocky Mountain spotted fever: a seasonal alert. Clin Infect Dis 20: 1111-1117.

    Article  CAS  PubMed  Google Scholar 

  • Walker DH, Cain BG, Olmstead PM (1978) Laboratory diagnosis of Rocky Mountain spotted fever by immunofluorescent demonstration of Rickettsia in Cutaneous lesions. Am J Clin Pathol 69: 619-623.

    Article  CAS  PubMed  Google Scholar 

  • Walker DH, Feng HM, Ladner S, Billings AN, Zaki SR, et al. (1997) Immunohistochemical diagnosis of typhus rickettsioses using an anti-lipopolysaccharide monoclonal antibody. Mod Pathol 10: 1038-1042.

    CAS  PubMed  Google Scholar 

  • Walker DH, Paddock CD, Dumler JS (2008) Emerging and re-emerging tick-transmitted rickettsial and ehrlichial infections. Med Clin North Am 92: 1345-1361, x.

    Google Scholar 

  • Walker DH, Parks FM, Betz TG, Taylor JP, Muehlberger JW (1989) Histopathology and immunohistologic demonstration of the distribution of Rickettsia typhi in fatal murine typhus. Am J Clin Pathol 91: 720-724.

    Article  CAS  PubMed  Google Scholar 

  • Walls JJ, Aguero-Rosenfeld M, Bakken JS, Goodman JL, Hossain D, et al. (1999) Inter- and intralaboratory comparison of Ehrlichia equi and human granulocytic ehrlichiosis (HGE) agent strains for serodiagnosis of HGE by the immunofluorescent-antibody test. J Clin Microbiol 37: 2968-2973.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Walter G, Botelho-Nevers E, Socolovschi C, Raoult D, Parola P (2012) Murine typhus in returned travelers: a report of thirty-two cases. Am J Trop Med Hyg 86: 1049-1053.

    Article  PubMed  PubMed Central  Google Scholar 

  • Wang X, Rikihisa Y, Lai TH, Kumagai Y, Zhi N, et al. (2004) Rapid sequential changeover of expressed p44 genes during the acute phase of Anaplasma phagocytophilum infection in horses. Infect Immun 72: 6852-6859.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yamamoto S, Minamishima Y (1982) Serodiagnosis of tsutsugamushi fever (scrub typhus) by the indirect immunoperoxidase technique. J Clin Microbiol 15: 1128-1132.

    CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Infectious Diseases, Department of Internal Medicine, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555-0435, USA

    Lucas S. Blanton

Authors
  1. Lucas S. Blanton
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lucas S. Blanton .

Editor information

Editors and Affiliations

  1. Lankenau Institute for Medical Research, Wynnewood, Pennsylvania, USA

    Sunil Thomas

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer International Publishing AG

About this chapter

Cite this chapter

Blanton, L.S. (2016). Rickettsiales: Laboratory Diagnosis. In: Thomas, S. (eds) Rickettsiales. Springer, Cham. https://doi.org/10.1007/978-3-319-46859-4_4

Download citation

Publish with us

Policies and ethics

Search

Navigation