Please use this identifier to cite or link to this item: http://hdl.handle.net/2445/171611
Title: Development of a multiplex real-time PCR surveillance assay for monitoring the health status of Ecuadorian amphibians at risk of extinction
Author: Burgos, Germán
Narváez-Narváez, David
Freire-Paspuel, Byron
Merino-Viteri, Andrés
Muslin, Claire
Genoy-Puerto, Alexander
Keywords: Amfibis
Extinció (Biologia)
Invertebrats
Amphibians
Extintion (Biology)
Invertebrates
Issue Date: 18-Oct-2019
Publisher: Elsevier B.V.
Abstract: Chytrid fungi and viruses within the genus Ranavirus have been associated with mass mortality events and declines in amphibian populations worldwide. The fungus Batrachochytrium dendrobatidis (Bd) was reported in Ecuador; however, other chytrid fungi like Batrachochytrium salamandrivorans (Bsal) or ranaviruses have not been described in the country so far. To prevent the introduction of pathogens into amphibian populations under conservation programs and to implement a successful disease surveillance program, the development of a sensitive and specific diagnostic assay was required. We describe here the optimization of one TaqMan probe-based multiplex quantitative polymerase chain reaction (qPCR) assay that enables the simultaneous detection of Bsal and ranavirus, and one monoplex TaqMan qPCR assay for the detection of Bd. Standard curves, with a high linear correlation (r2 > 0.995), were generated using a synthetic genome template (gBlocks®) containing the target sequences from all three pathogens. Different samples from skin, liver, kidney, spleen, and lung from six different amphibian species were tested, and both qPCR assays showed highly reproducible and reliable results. To our knowledge, this method is the first multiplex qPCR system developed in Ecuador for identifying amphibian pathogens and represents a valuable tool for the early detection of these pathogens and for infection and co-infection monitoring in future epidemiological surveillance of amphibian species at risk of extinction.
Note: Versió postprint del document publicat a: https://doi.org/10.1016/j.fsigss.2019.10.134
It is part of: Forensic Science International: Genetics Supplement Series, 2019, vol. 7, num. 1, p. 674-676
URI: http://hdl.handle.net/2445/171611
Related resource: https://doi.org/10.1016/j.fsigss.2019.10.134
ISSN: 1875-1768
Appears in Collections:Articles publicats en revistes (Farmàcia, Tecnologia Farmacèutica i Fisicoquímica)

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