Development of a Quantitative PCR Assay for Differentiating the Agent of Heartwater Disease, Ehrlichia ruminantium, from the Panola Mountain Ehrlichia

Summary Panola Mountain Ehrlichia (PME) is an emerging Ehrlichia sp. reported in ten US states. Based on the sequence homology of all known genes, PME is closely related to Ehrlichia ruminantium (ER), the causative agent of heartwater. Heartwater is an economically important tick‐borne disease of ca...

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Bibliographic Details
Published in:Transboundary and emerging diseases Vol. 63; no. 6; pp. e260 - e269
Main Authors: Sayler, K. A., Loftis, A. D., Mahan, S. M., Barbet, A. F.
Format: Journal Article
Language:English
Published: Germany Blackwell Publishing Ltd 01.12.2016
John Wiley & Sons, Inc
Subjects:
Africa
Animal diseases
Animals
Caribbean
Cattle
Cattle Diseases - epidemiology
cross reaction
diagnostic qPCR
disease surveillance
DNA
DNA Primers
Economic importance
Ehrlichia - genetics
Ehrlichia - isolation & purification
Ehrlichia ruminantium
Ehrlichia ruminantium - genetics
Ehrlichia ruminantium - isolation & purification
emerging diseases
Enzyme-Linked Immunosorbent Assay
epidemiological studies
Epidemiology
foreign animal disease
genes
Goats
heartwater
Heartwater Disease - diagnosis
Heartwater Disease - epidemiology
islands
Livestock
livestock losses
Outbreaks
Panola Mountain Ehrlichia
pathogens
Peptides
Polymerase chain reaction
quantitative polymerase chain reaction
Real-Time Polymerase Chain Reaction
sequence homology
Sheep
Sub-Saharan Africa
United States
United States - epidemiology
Caribbean
United States
Sub-Saharan Africa
heartwater
diagnostic qPCR
livestock losses
Panola Mountain Ehrlichia
foreign animal disease
Ehrlichia ruminantium
ISSN:1865-1674, 1865-1682
Online Access:Get full text
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Summary:Summary Panola Mountain Ehrlichia (PME) is an emerging Ehrlichia sp. reported in ten US states. Based on the sequence homology of all known genes, PME is closely related to Ehrlichia ruminantium (ER), the causative agent of heartwater. Heartwater is an economically important tick‐borne disease of cattle, sheep and goats responsible for stock losses in sub‐Saharan Africa. Unfortunately, ER was imported to the Caribbean islands in the 19th century, and the presence of this foreign animal disease in the Caribbean poses a threat to the US mainland. If introduced, a heartwater outbreak would cause massive losses of naïve livestock. The serologic assay of choice to diagnose heartwater is cross‐reactive with Ehrlichia spp., including PME, as we demonstrate here, which would confound disease surveillance in the event of a heartwater outbreak. The purpose of this study was to develop a diagnostic assay capable of rapidly distinguishing between these pathogens. Using synthetic MAP‐1B peptides for ER and PME, we tested the cross‐reactivity of this assay using sera from infected livestock. The MAP‐1B ELISA cannot distinguish between animals infected with PME and ER. Therefore, a dual‐plex Taqman™ qPCR assay targeting the groEL gene of PME and ER was developed and validated. Primers were designed that are conserved among all known strains of ER, allowing for the amplification of strains from the Caribbean and Africa. The assay is highly sensitive (10 copies of DNA) and specific. This assay distinguishes between infection with PME and ER and will be a valuable tool in the event of heartwater outbreak on the US mainland, or for epidemiological studies involving either disease‐causing organism.
Bibliography:istex:582AA852318B5D22DA7CE68266A35AFA81C19162
ArticleID:TBED12339
ark:/67375/WNG-TJS41VRG-9
U.S. Department of Agriculture - No. 2010-34135-21053
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SourceType-Scholarly Journals-1
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ISSN:1865-1674
1865-1682
DOI:10.1111/tbed.12339