An indirect immunofluorescence assay using a cell culture-derived antigen for detection of antibodies to the agent of human granulocytic ehrlichiosis.

Journal of Clinical Microbiology
W L NicholsonJames E Childs

Abstract

An indirect immunofluorescence assay for the detection of human antibodies to the agent of human granulocytic ehrlichiosis (HGE) was developed and standardized. Antigen was prepared from a human promyelocytic leukemia cell line (HL-60) infected with a tick-derived isolate of the HGE agent (USG3). Suitable antigen presentation and preservation of cellular morphology were obtained when infected cells were applied and cultured on the slide, excess medium was removed, and cells were fixed with acetone. Use of a buffer containing bovine serum albumin and goat serum reduced background fluorescence, and use of an immunoglobulin G (gamma-specific) conjugate reduced nonspecific binding. The assay readily detected specific antibody from HGE patients and did not detect antibody from healthy individuals. No significant reactivity was noted in sera from patients with high titers of antibodies to other rickettsial species. We were able to identify antibodies reactive to USG3 antigen in samples from areas where HGE is endemic that had tested negative to other rickettsial agents. Animal sera reactive against Ehrlichia equi or Ehrlichia phagocytophila bound to the HGE antigen, indicating that the assay may be useful for veterinary use. Comparab...Continue Reading

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Citations

Oct 8, 1999·Emerging Infectious Diseases·J H McQuistonJ E Childs
Jun 14, 2003·Vector Borne and Zoonotic Diseases·Maria E Aguero-Rosenfeld
Aug 6, 2014·The American Journal of Tropical Medicine and Hygiene·Jennifer H McQuistonJohn Dunn
Jun 5, 1998·Annual Review of Public Health·D H Walker
Aug 31, 2001·Clinical and Diagnostic Laboratory Immunology·S Lotric-FurlanF Strle
Sep 7, 2002·DNA and Cell Biology·Robert F MassungAsiya Gusa
Jul 8, 2000·Journal of Internal Medicine·N PickI Schwartz
Jul 16, 2003·Annals of the New York Academy of Sciences·William L NicholsonJames E Childs
Dec 1, 2006·Annals of the New York Academy of Sciences·Eugenio LilliniManuela Scarpulla
Mar 6, 2010·Emerging Infectious Diseases·Jennifer AdjemianJeffery Taylor
May 8, 2013·Veterinary Clinical Pathology·Robin W Allison, Susan E Little
Dec 4, 2001·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Johan S BakkenJ Stephen Dumler
Jan 4, 2001·Clinical and Diagnostic Laboratory Immunology·P BrouquiD Raoult
May 2, 2008·Journal of Medical Case Reports·Will K ReevesAlan G Czarkowski
Jul 28, 2012·Vector Borne and Zoonotic Diseases·Jennifer AdjemianDavid Wong
Jul 20, 2001·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·S Lotric-FurlanF Strle
Sep 20, 2006·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·Charmaine Leung-Shea, Patrick J Danaher
Nov 23, 2006·Annals of the New York Academy of Sciences·Robert F MassungTimothy J Kurtti
May 17, 2011·Acta Veterinaria Scandinavica·Lise GrøvaSnorre Stuen
Aug 5, 2011·The New England Journal of Medicine·Bobbi S PrittMarina E Eremeeva
Sep 15, 2000·Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America·J S Bakken, J S Dumler
Jul 16, 2003·Annals of the New York Academy of Sciences·Stanka Lotric-FurlanFranc Strle
May 4, 2007·Journal of Clinical Microbiology·Robert F MassungTimothy J Kurtti
Aug 2, 2000·Journal of Clinical Microbiology·R F MassungC I Murphy
Feb 8, 2003·Journal of Clinical Microbiology·Robert F Massung, Kimetha G Slater

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