Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks

Background: Ehrlichia and Rickettsia are two major rickettsial genera transmitted by ticks that affect a number of wild and domestic animal species and human populations around the world. Objective: To design and validate a duplex PCR for Ehrlichia and Rickettsia in ticks. Methods: Assay validation...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Juan C. Pérez Pérez, Carolina Montoya Ruiz, Esteban Arroyave Sierra, Luis E. Paternina, Juan D. Rodas
Formato:	article
Lenguaje:	EN
Publicado:	Universidad de Antioquia 2018
Materias:	exclusiveness groel inclusiveness robustness rrna 23s Animal culture SF1-1100
Acceso en línea:	https://doaj.org/article/2f1d6e25f23c46919f913fcb31cc42cc
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:2f1d6e25f23c46919f913fcb31cc42cc
record_format	dspace
spelling	oai:doaj.org-article:2f1d6e25f23c46919f913fcb31cc42cc2021-12-01T19:35:11ZDesign and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks2256-295810.17533/udea.rccp.v31n4a05https://doaj.org/article/2f1d6e25f23c46919f913fcb31cc42cc2018-12-01T00:00:00Zhttps://revistas.udea.edu.co/index.php/rccp/article/view/330532https://doaj.org/toc/2256-2958Background: Ehrlichia and Rickettsia are two major rickettsial genera transmitted by ticks that affect a number of wild and domestic animal species and human populations around the world. Objective: To design and validate a duplex PCR for Ehrlichia and Rickettsia in ticks. Methods: Assay validation included testing for sensitivity, specificity, reproducibility, and robustness of the PCR. The groEL and 23sr RNA genes were used for Ehrlichia and Rickettsia, respectively. Results: The limit of detection was one hundred gene copies per 50 μL of reaction for Ehrlichia spp, and one gene copy of Rickettsia per 50 μL of reaction. In general, the primers of the test only amplified in silico those bacterial agents for which they were originally designed, with the exception of the primers for Rickettsia that also amplified Methylocystis sp. The test was reproducible (intermediate precision) 96.7% of the times for both agents. The test was robust enough to tolerate concentration changes of all reagents with the exception of Taq DNA polymerase. Conclusions: The validation results indicated that this PCR is useful for detection in both bacterial genera and it is a good candidate for diagnostic validation.Juan C. Pérez PérezCarolina Montoya RuizEsteban Arroyave SierraLuis E. PaterninaJuan D. RodasUniversidad de Antioquiaarticleexclusivenessgroelinclusivenessrobustnessrrna 23sAnimal cultureSF1-1100ENRevista Colombiana de Ciencias Pecuarias, Vol 31, Iss 4, Pp 285-294 (2018)
institution	DOAJ
collection	DOAJ
language	EN
topic	exclusiveness groel inclusiveness robustness rrna 23s Animal culture SF1-1100
spellingShingle	exclusiveness groel inclusiveness robustness rrna 23s Animal culture SF1-1100 Juan C. Pérez Pérez Carolina Montoya Ruiz Esteban Arroyave Sierra Luis E. Paternina Juan D. Rodas Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks
description	Background: Ehrlichia and Rickettsia are two major rickettsial genera transmitted by ticks that affect a number of wild and domestic animal species and human populations around the world. Objective: To design and validate a duplex PCR for Ehrlichia and Rickettsia in ticks. Methods: Assay validation included testing for sensitivity, specificity, reproducibility, and robustness of the PCR. The groEL and 23sr RNA genes were used for Ehrlichia and Rickettsia, respectively. Results: The limit of detection was one hundred gene copies per 50 μL of reaction for Ehrlichia spp, and one gene copy of Rickettsia per 50 μL of reaction. In general, the primers of the test only amplified in silico those bacterial agents for which they were originally designed, with the exception of the primers for Rickettsia that also amplified Methylocystis sp. The test was reproducible (intermediate precision) 96.7% of the times for both agents. The test was robust enough to tolerate concentration changes of all reagents with the exception of Taq DNA polymerase. Conclusions: The validation results indicated that this PCR is useful for detection in both bacterial genera and it is a good candidate for diagnostic validation.
format	article
author	Juan C. Pérez Pérez Carolina Montoya Ruiz Esteban Arroyave Sierra Luis E. Paternina Juan D. Rodas
author_facet	Juan C. Pérez Pérez Carolina Montoya Ruiz Esteban Arroyave Sierra Luis E. Paternina Juan D. Rodas
author_sort	Juan C. Pérez Pérez
title	Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks
title_short	Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks
title_full	Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks
title_fullStr	Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks
title_full_unstemmed	Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks
title_sort	design and analytical validation of a duplex pcr for ehrlichia and rickettsia detection in ticks
publisher	Universidad de Antioquia
publishDate	2018
url	https://doaj.org/article/2f1d6e25f23c46919f913fcb31cc42cc
work_keys_str_mv	AT juancperezperez designandanalyticalvalidationofaduplexpcrforehrlichiaandrickettsiadetectioninticks AT carolinamontoyaruiz designandanalyticalvalidationofaduplexpcrforehrlichiaandrickettsiadetectioninticks AT estebanarroyavesierra designandanalyticalvalidationofaduplexpcrforehrlichiaandrickettsiadetectioninticks AT luisepaternina designandanalyticalvalidationofaduplexpcrforehrlichiaandrickettsiadetectioninticks AT juandrodas designandanalyticalvalidationofaduplexpcrforehrlichiaandrickettsiadetectioninticks
_version_	1718404621814202368

Design and analytical validation of a duplex PCR for Ehrlichia and Rickettsia detection in ticks

Ejemplares similares