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Pooling skin swabs does not inhibit qPCR detection of amphibian chytrid infection
- Source :
- PLoS ONE, Vol 14, Iss 4, p e0214405 (2019), PLoS ONE 14(4): e0214405. https://doi.org/10.1371/journal.pone.0214405--PLoS ONE--http://www.bibliothek.uni-regensburg.de/ezeit/?2267670--http://www.plosone.org/home.action--https://journals.plos.org/plosone/--https://www.ncbi.nlm.nih.gov/pmc/journals/440/--1932-6203--1932-6203, PLOS ONE, PLoS ONE
- Publication Year :
- 2019
- Publisher :
- Public Library of Science (PLoS), 2019.
-
Abstract
- Immediate and reliable pathogen detection in large numbers of samples is essential in wildlife disease monitoring and is often realized by DNA-based techniques. Pooling samples increases processing efficiency and reduces processing costs, and has been suggested as a viable technique for quantitative PCR detection of fungal amphibian pathogens of the genus Batrachochytrium. For these fungi, this diagnostic method has been validated by in vitro set ups that provided controlled test conditions but did not take into account potential effects from amphibian skin compounds (e.g. skin secretions and Microbiota) on the approach. Some of these skin compounds are known to cause PCR inhibition in single sample applications and could lead to false negative reactions and thereby hamper pathogen detection. In this study we examined the effect of skin compounds on the pooled extraction method by swabbing individuals of seven amphibian species (one Anura and six Caudata) prior to the inoculation of the swabs with chytrid zoospores. For each species, swabs were extracted in pools of different sizes (from one to four swabs) with only one swab per pool being inoculated with zoospores. There were no significant differences regarding the ability to detect zoospores when comparing pool sizes for any species, with a tendency for more false negatives when the inoculated swab had been inoculated with a single zoospore. This study provides further in vivo evidence for the viability of the pooled extraction method for DNA-based detection of pathogens.
- Subjects :
- Caudata
Science
Animals, Wild
Mycology
Pathology and Laboratory Medicine
Infections
Real-Time Polymerase Chain Reaction
Microbiology
Article
Specimen Handling
Amphibians
BATRACHOCHYTRIUM-SALAMANDRIVORANS
Extraction techniques
DENDROBATIDIS
Medicine and Health Sciences
Genetics
Animals
Veröffentlichung der TU Braunschweig
Veterinary Sciences
Microbial Pathogens
DNA extraction
Skin
ddc:5
Fungal Pathogens
Evolutionary Biology
Population Biology
Organisms
Fungi
Biology and Life Sciences
Eukaryota
Gene Pool
DNA
Research and analysis methods
ddc:57
Chytridiomycota
Veterinary Diseases
FUNGUS
Medical Microbiology
Vertebrates
SP-NOV
Medicine
Veterinary Science
Pathogens
Anura
Publikationsfonds der TU Braunschweig
Population Genetics
Research Article
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 14
- Issue :
- 4
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.pmid.dedup....62c0f3928d7eed767c95b0525d3d7fdb