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

Michelle E J Allerdice, Bobbi S. Pritt, Lynne M. Sloan, Christopher D. Paddock, Sandor E. Karpathy

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

The Ehrlichia muris-like agent (EMLA) is an emerging, tick-transmitted human pathogen that occurs in the upper Midwestern United States. Here, we describe the development and validation of a p13-based quantitative real-time PCR TaqMan assay to detect EMLA in blood or tissues of ticks, humans, and rodents. The primer and probe specificities of the assay were ascertained using a large panel of various Ehrlichia species and other members of Rickettsiales. In addition to control DNA, both non-infected and EMLA-infected human blood, Mus musculus blood, and M. musculus tissue extracts were evaluated, as were non-infected and EMLA-infected Ixodes scapularis and uninfected Dermacentor variabilis DNA lysates. The specificity of the probe was determined via real-time PCR. An EMLA p13 control plasmid was constructed, and serial dilutions were used to determine the analytical sensitivity, which was found to be 1 copy per 4. μl of template DNA. The sensitivity and specificity of this assay provides a powerful tool for ecological studies involving arthropod vectors and their mammalian hosts.

Original languageEnglish (US)
Pages (from-to)146-149
Number of pages4
JournalTicks and Tick-borne Diseases
Volume7
Issue number1
DOIs
StatePublished - Feb 1 2016

Keywords

  • Ehrlichia muris
  • Ehrlichia muris-like (EML) agent
  • Ehrlichiosis
  • Tick-borne disease

ASJC Scopus subject areas

  • Infectious Diseases
  • Insect Science
  • Parasitology
  • Microbiology

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