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

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

doi:10.1016/j.ttbdis.2015.10.004

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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

Published Date:

Oct 22 2015

Publisher's site:

http://dx.doi.org/10.1016/j.ttbdis.2015.10.004 New Site Icon

Source:

Ticks Tick Borne Dis. 7(1):146-149.

[PDF-284.76 KB]

Details:

Personal Authors:

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

Keywords:

[+]

Pubmed ID:

26507653

Pubmed Central ID:

PMC4659714

Description:

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 4μl of template DNA. The sensitivity and specificity of this assay provides a powerful tool for ecological studies involving arthropod vectors and their mammalian hosts.

Document Type:

Journal Article

Collection(s):

CDC Public Access

Funding:

CC999999/Intramural CDC HHS/United States

Supporting Files:

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