TY - JOUR
T1 - 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
AU - Allerdice, Michelle E.J.
AU - Pritt, Bobbi S.
AU - Sloan, Lynne M.
AU - Paddock, Christopher D.
AU - Karpathy, Sandor E.
N1 - Publisher Copyright:
© 2015 .
PY - 2016/2
Y1 - 2016/2
N2 - 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.
AB - 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.
KW - Ehrlichia muris
KW - Ehrlichia muris-like (EML) agent
KW - Ehrlichiosis
KW - Tick-borne disease
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UR - http://www.scopus.com/inward/citedby.url?scp=84947447965&partnerID=8YFLogxK
U2 - 10.1016/j.ttbdis.2015.10.004
DO - 10.1016/j.ttbdis.2015.10.004
M3 - Article
C2 - 26507653
AN - SCOPUS:84947447965
SN - 1877-959X
VL - 7
SP - 146
EP - 149
JO - Ticks and Tick-borne Diseases
JF - Ticks and Tick-borne Diseases
IS - 1
ER -