Evaluation of an improved PCR diagnostic assay for human granulocytic ehrlichiosis

Daniel C. Edelman, J. Stephen Dumler

Research output: Contribution to journalArticle

Abstract

Background: Human granulocytic ehrlichiosis (HGE) is a tick-borne disease caused by an agent similar to Ehrlichia equi. The etiologic agent has only recently been cultivated, and clinical and laboratory findings are nonspecific. Severity and fatal outcome are associated with delays in diagnosis and therapy. Methods and Results: The sensitivity of the polymerase chain reaction (PCR) technique for HGE was enhanced at least 250-fold for identification of the 16S ribosomal RNA gene of the HGE agent in blood by shortening cycle segments, using 35 rounds of amplification, employing eLONGase polymerase, and by using SYBR Green I for detection of amplified products. From 17 patients with suspected HGE, acute-phase blood was tested by PCR and for E. equi antibodies in acute and convalescent sera. The HGE PCR detected ehrlichial DNA in 1 ng of genomic leukocyte DNA and in as little as 0.6 infected leukocyte per microliter of blood. Of the 17 patients with suspected HGE, 7 (41%) were serolpgically confirmed, and six of these seven had positive PCRs (86% sensitivity). Ten seronegative patients had negative PCRs, and no patient had a positive PCR and negative serology (100% specificity). Conclusion: The polymerase chain reaction for the HGE agent 16S rRNA gene is a sensitive and specific method for the early confirmation of HGE at a time when therapeutic decisions are required.

Original languageEnglish (US)
Pages (from-to)41-49
Number of pages9
JournalMolecular Diagnosis
Volume1
Issue number1
StatePublished - 1996

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Ehrlichiosis
Polymerase Chain Reaction
Anaplasma phagocytophilum
rRNA Genes
Leukocytes
Tick-Borne Diseases
16S Ribosomal RNA
Fatal Outcome
DNA
Serology
Antibodies

Keywords

  • Ehrlichia equi
  • Human granulocytic ehrlichiosis
  • Sybr green I
  • Tick-borne disease

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Evaluation of an improved PCR diagnostic assay for human granulocytic ehrlichiosis. / Edelman, Daniel C.; Stephen Dumler, J.

In: Molecular Diagnosis, Vol. 1, No. 1, 1996, p. 41-49.

Research output: Contribution to journalArticle

Edelman, DC & Stephen Dumler, J 1996, 'Evaluation of an improved PCR diagnostic assay for human granulocytic ehrlichiosis', Molecular Diagnosis, vol. 1, no. 1, pp. 41-49.
Edelman, Daniel C. ; Stephen Dumler, J. / Evaluation of an improved PCR diagnostic assay for human granulocytic ehrlichiosis. In: Molecular Diagnosis. 1996 ; Vol. 1, No. 1. pp. 41-49.
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