Development of a Multiplex Droplet Digital PCR Assay for the Detection of Babesia, Bartonella, and Borrelia Species

We describe the development, optimization, and validation of a multiplex droplet digital PCR (ddPCR) assay for the simultaneous detection of Babesia, Bartonella, and Borrelia spp. DNA from several sample matrices, including clinical blood samp...

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Autores principales:	Ricardo Maggi, Edward B. Breitschwerdt, Barbara Qurollo, Jennifer C. Miller
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	<i>Babesia</i> <i>Bartonella</i> <i>Borrelia</i> <i>Cytauxzoon</i> Piroplasma <i>Theileria</i> Medicine R
Acceso en línea:	https://doaj.org/article/7b2d1c3dd197434f9569ef23d15bdc74
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spelling	oai:doaj.org-article:7b2d1c3dd197434f9569ef23d15bdc742021-11-25T18:38:26ZDevelopment of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species10.3390/pathogens101114622076-0817https://doaj.org/article/7b2d1c3dd197434f9569ef23d15bdc742021-11-01T00:00:00Zhttps://www.mdpi.com/2076-0817/10/11/1462https://doaj.org/toc/2076-0817We describe the development, optimization, and validation of a multiplex droplet digital PCR (ddPCR) assay for the simultaneous detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> spp. DNA from several sample matrices, including clinical blood samples from animals and humans, vectors, in-vitro infected human and animal cell lines, and tissues obtained from animal models (infected with <i>Bartonella</i> and/or <i>B. burgdorferi</i>). The multiplex ddPCR assay was able to detect 31 <i>Bartonella</i>, 13 <i>Borrelia</i>, and 24 <i>Babesia</i> species, including <i>Theileria equi</i>, <i>T. cervi</i>, and <i>Cytauxzoon felis</i>. No amplification of <i>Treponema</i> or <i>Leptospira</i> spp. was observed. Sensitivity of 0.2–5 genome equivalent DNA copies per microliter was achieved for different members of the <i>Bartonella</i> and Borrelia genus, depending on the species or matrix type (water or spiked blood DNA) tested. The ddPCR assay facilitated the simultaneous detection of co-infections with two and three vector-borne pathogens comprising four different genera (<i>Babesia</i>, <i>Bartonella</i>, <i>Borrelia</i>, and <i>Theileria</i>) from clinical and other sample sources.Ricardo MaggiEdward B. BreitschwerdtBarbara QurolloJennifer C. MillerMDPI AGarticle<i>Babesia</i><i>Bartonella</i><i>Borrelia</i><i>Cytauxzoon</i>Piroplasma<i>Theileria</i>MedicineRENPathogens, Vol 10, Iss 1462, p 1462 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	<i>Babesia</i> <i>Bartonella</i> <i>Borrelia</i> <i>Cytauxzoon</i> Piroplasma <i>Theileria</i> Medicine R
spellingShingle	<i>Babesia</i> <i>Bartonella</i> <i>Borrelia</i> <i>Cytauxzoon</i> Piroplasma <i>Theileria</i> Medicine R Ricardo Maggi Edward B. Breitschwerdt Barbara Qurollo Jennifer C. Miller Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species
description	We describe the development, optimization, and validation of a multiplex droplet digital PCR (ddPCR) assay for the simultaneous detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> spp. DNA from several sample matrices, including clinical blood samples from animals and humans, vectors, in-vitro infected human and animal cell lines, and tissues obtained from animal models (infected with <i>Bartonella</i> and/or <i>B. burgdorferi</i>). The multiplex ddPCR assay was able to detect 31 <i>Bartonella</i>, 13 <i>Borrelia</i>, and 24 <i>Babesia</i> species, including <i>Theileria equi</i>, <i>T. cervi</i>, and <i>Cytauxzoon felis</i>. No amplification of <i>Treponema</i> or <i>Leptospira</i> spp. was observed. Sensitivity of 0.2–5 genome equivalent DNA copies per microliter was achieved for different members of the <i>Bartonella</i> and Borrelia genus, depending on the species or matrix type (water or spiked blood DNA) tested. The ddPCR assay facilitated the simultaneous detection of co-infections with two and three vector-borne pathogens comprising four different genera (<i>Babesia</i>, <i>Bartonella</i>, <i>Borrelia</i>, and <i>Theileria</i>) from clinical and other sample sources.
format	article
author	Ricardo Maggi Edward B. Breitschwerdt Barbara Qurollo Jennifer C. Miller
author_facet	Ricardo Maggi Edward B. Breitschwerdt Barbara Qurollo Jennifer C. Miller
author_sort	Ricardo Maggi
title	Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species
title_short	Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species
title_full	Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species
title_fullStr	Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species
title_full_unstemmed	Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species
title_sort	development of a multiplex droplet digital pcr assay for the detection of <i>babesia</i>, <i>bartonella</i>, and <i>borrelia</i> species
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/7b2d1c3dd197434f9569ef23d15bdc74
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Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species

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