A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for Anaplasma/Ehrlichia in Roaming Dogs and Symptomatic Dogs in Trinidad

This study compared two methods to detect cases of canine ehrlichiosis in a field setting. One method was a polymerase chain reaction for the 16S rRNA gene followed by reverse line blot hybridisation with genera and species-specific probes for Anaplasma/Ehrlichia. The second metho...

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Autores principales:	Karla Georges, Chuckwudozi Ezeokoli, Godwin Isitor, Alex Mutani, Olivier Sparagano, Candice Sant
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	cell culture dogs <i>Ehrlichia canis</i> polymerase chain reaction reverse line blot hybridisation Trinidad Medicine R
Acceso en línea:	https://doaj.org/article/b0701ee8fba94d6baa44062640b8686c
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spelling	oai:doaj.org-article:b0701ee8fba94d6baa44062640b8686c2021-11-25T18:38:13ZA Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad10.3390/pathogens101114312076-0817https://doaj.org/article/b0701ee8fba94d6baa44062640b8686c2021-11-01T00:00:00Zhttps://www.mdpi.com/2076-0817/10/11/1431https://doaj.org/toc/2076-0817This study compared two methods to detect cases of canine ehrlichiosis in a field setting. One method was a polymerase chain reaction for the 16S rRNA gene followed by reverse line blot hybridisation with genera and species-specific probes for <i>Anaplasma/Ehrlichia</i>. The second method was an autologous cell culture of peripheral leucocytes isolated from heparinised blood and maintained in a homologous canine serum in Dulbecco’s Modified Eagle medium without antibiotics. The cultures were examined under light microscopy for inclusion bodies after 48 h. Leucocytes were successfully propagated for 20 of the 34 samples submitted for autologous cell culture. Inclusion bodies were observed after cell culture in leucocytes of eight dogs. Two dogs were positive to the <i>Anaplasma/Ehrlichia</i> genera probe and six dogs were positive to the <i>E. canis</i> probe after reverse line blot hybridisation. There was acceptable agreement between reverse line blot hybridisation and cell culture results. Both reverse line blot hybridisation and autologous cell cultures can be used to detect <i>E. canis</i> in subclinical and clinical cases of disease. A definitive diagnosis of <i>E. canis</i> is best achieved by a combination of clinical signs, positive autologous cell culture, and reverse line blot hybridisation results.Karla GeorgesChuckwudozi EzeokoliGodwin IsitorAlex MutaniOlivier SparaganoCandice SantMDPI AGarticlecell culturedogs<i>Ehrlichia canis</i>polymerase chain reactionreverse line blot hybridisationTrinidadMedicineRENPathogens, Vol 10, Iss 1431, p 1431 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cell culture dogs <i>Ehrlichia canis</i> polymerase chain reaction reverse line blot hybridisation Trinidad Medicine R
spellingShingle	cell culture dogs <i>Ehrlichia canis</i> polymerase chain reaction reverse line blot hybridisation Trinidad Medicine R Karla Georges Chuckwudozi Ezeokoli Godwin Isitor Alex Mutani Olivier Sparagano Candice Sant A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad
description	This study compared two methods to detect cases of canine ehrlichiosis in a field setting. One method was a polymerase chain reaction for the 16S rRNA gene followed by reverse line blot hybridisation with genera and species-specific probes for <i>Anaplasma/Ehrlichia</i>. The second method was an autologous cell culture of peripheral leucocytes isolated from heparinised blood and maintained in a homologous canine serum in Dulbecco’s Modified Eagle medium without antibiotics. The cultures were examined under light microscopy for inclusion bodies after 48 h. Leucocytes were successfully propagated for 20 of the 34 samples submitted for autologous cell culture. Inclusion bodies were observed after cell culture in leucocytes of eight dogs. Two dogs were positive to the <i>Anaplasma/Ehrlichia</i> genera probe and six dogs were positive to the <i>E. canis</i> probe after reverse line blot hybridisation. There was acceptable agreement between reverse line blot hybridisation and cell culture results. Both reverse line blot hybridisation and autologous cell cultures can be used to detect <i>E. canis</i> in subclinical and clinical cases of disease. A definitive diagnosis of <i>E. canis</i> is best achieved by a combination of clinical signs, positive autologous cell culture, and reverse line blot hybridisation results.
format	article
author	Karla Georges Chuckwudozi Ezeokoli Godwin Isitor Alex Mutani Olivier Sparagano Candice Sant
author_facet	Karla Georges Chuckwudozi Ezeokoli Godwin Isitor Alex Mutani Olivier Sparagano Candice Sant
author_sort	Karla Georges
title	A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad
title_short	A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad
title_full	A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad
title_fullStr	A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad
title_full_unstemmed	A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad
title_sort	comparison of peripheral blood smears, autologous cell cultures, and reverse line blot hybridisation in screening for <i>anaplasma</i>/<i>ehrlichia</i> in roaming dogs and symptomatic dogs in trinidad
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/b0701ee8fba94d6baa44062640b8686c
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A Comparison of Peripheral Blood Smears, Autologous Cell Cultures, and Reverse Line Blot Hybridisation in Screening for <i>Anaplasma</i>/<i>Ehrlichia</i> in Roaming Dogs and Symptomatic Dogs in Trinidad

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