Development of a Multiplex Droplet Digital PCR Assay for the Detection of <i>Babesia</i>, <i>Bartonella</i>, and <i>Borrelia</i> Species
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 samp...
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| Autores principales: | , , , |
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| Formato: | article |
| Lenguaje: | EN |
| Publicado: |
MDPI AG
2021
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| Materias: | |
| Acceso en línea: | https://doaj.org/article/7b2d1c3dd197434f9569ef23d15bdc74 |
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| Sumario: | 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. |
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