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Pooling skin swabs does not inhibit qPCR detection of amphibian chytrid infection

(2019) PLOS ONE. 14(4).
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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.
Keywords
BATRACHOCHYTRIUM-SALAMANDRIVORANS, SP-NOV, DENDROBATIDIS, FUNGUS

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Citation

Please use this url to cite or link to this publication:

MLA
Sabino-Pinto, Joana et al. “Pooling Skin Swabs Does Not Inhibit qPCR Detection of Amphibian Chytrid Infection.” PLOS ONE 14.4 (2019): n. pag. Print.
APA
Sabino-Pinto, J., Martel, A., Pasmans, F., Steinfartz, S., & Vences, M. (2019). Pooling skin swabs does not inhibit qPCR detection of amphibian chytrid infection. PLOS ONE, 14(4).
Chicago author-date
Sabino-Pinto, Joana, An Martel, Frank Pasmans, Sebastian Steinfartz, and Miguel Vences. 2019. “Pooling Skin Swabs Does Not Inhibit qPCR Detection of Amphibian Chytrid Infection.” Plos One 14 (4).
Chicago author-date (all authors)
Sabino-Pinto, Joana, An Martel, Frank Pasmans, Sebastian Steinfartz, and Miguel Vences. 2019. “Pooling Skin Swabs Does Not Inhibit qPCR Detection of Amphibian Chytrid Infection.” Plos One 14 (4).
Vancouver
1.
Sabino-Pinto J, Martel A, Pasmans F, Steinfartz S, Vences M. Pooling skin swabs does not inhibit qPCR detection of amphibian chytrid infection. PLOS ONE. 2019;14(4).
IEEE
[1]
J. Sabino-Pinto, A. Martel, F. Pasmans, S. Steinfartz, and M. Vences, “Pooling skin swabs does not inhibit qPCR detection of amphibian chytrid infection,” PLOS ONE, vol. 14, no. 4, 2019.
@article{8625945,
  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.},
  articleno    = {e0214405},
  author       = {Sabino-Pinto, Joana and Martel, An and Pasmans, Frank and Steinfartz, Sebastian and Vences, Miguel},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keywords     = {BATRACHOCHYTRIUM-SALAMANDRIVORANS,SP-NOV,DENDROBATIDIS,FUNGUS},
  language     = {eng},
  number       = {4},
  pages        = {9},
  title        = {Pooling skin swabs does not inhibit qPCR detection of amphibian chytrid infection},
  url          = {http://dx.doi.org/10.1371/journal.pone.0214405},
  volume       = {14},
  year         = {2019},
}

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