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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Vydané v:	Transboundary and emerging diseases Ročník 63; číslo 6; s. e260 - e269
Hlavní autori:	Sayler, K. A., Loftis, A. D., Mahan, S. M., Barbet, A. F.
Médium:	Journal Article
Jazyk:	English
Vydavateľské údaje:	Germany Blackwell Publishing Ltd 01.12.2016 John Wiley & Sons, Inc
Predmet:	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
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Shrnutí:	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.
Bibliografia:	istex:582AA852318B5D22DA7CE68266A35AFA81C19162 ArticleID:TBED12339 ark:/67375/WNG-TJS41VRG-9 U.S. Department of Agriculture - No. 2010-34135-21053 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23
ISSN:	1865-1674 1865-1682
DOI:	10.1111/tbed.12339