PUBLICATION
Detection of the parasitic nematode, Pseudocapillaria tomentosa, in zebrafish tissues and environmental DNA in research aquaria
- Authors
- Norris, L., Lawler, N., Hunkapiller, A., Mulrooney, D.M., Kent, M.L., Sanders, J.L.
- ID
- ZDB-PUB-200729-5
- Date
- 2020
- Source
- Journal of fish diseases 43(9): 1087-1095 (Journal)
- Registered Authors
- Kent, Michael
- Keywords
- Pseudocapillaria, SSU-rDNA, eDNA, real-time PCR, zebrafish
- MeSH Terms
-
- Animals
- DNA, Environmental/analysis*
- DNA, Helminth/analysis
- Fish Diseases/diagnosis
- Fish Diseases/parasitology*
- Laboratory Animal Science
- Nematoda/genetics
- Nematoda/isolation & purification*
- Nematode Infections/diagnosis
- Nematode Infections/veterinary
- Real-Time Polymerase Chain Reaction/methods
- Zebrafish*
- PubMed
- 32720361 Full text @ J. Fish Dis.
Citation
Norris, L., Lawler, N., Hunkapiller, A., Mulrooney, D.M., Kent, M.L., Sanders, J.L. (2020) Detection of the parasitic nematode, Pseudocapillaria tomentosa, in zebrafish tissues and environmental DNA in research aquaria. Journal of fish diseases. 43(9):1087-1095.
Abstract
Although zebrafish continue to increase in popularity as a vertebrate animal model for biomedical research, chronic infectious diseases in laboratory populations remain prevalent. The presence of pathogens such as Pseudocapillaria tomentosa, a parasitic nematode found in the intestine of infected zebrafish, can significantly influence experimental endpoints and negatively impact reproducibility of research findings. Thus, there is a need for screening tests for zebrafish with the sensitivity to detect even low levels of pathogens present in tissues. Assays based on the detection of DNA are commonly used for such screening tests. Newer technologies such as digital PCR provide an opportunity to improve the sensitivity and precision of these assays, so they can be reliably used to detect pathogen DNA in water, reducing the need for lethal testing. We have designed a qPCR-based assay with the sensitivity to detect less than 5 copies of the P. tomentosa SSU-rDNA gene in tissues of infected zebrafish and environmental DNA from aquarium water housing infected fish. In addition, we adapted this test to a dPCR platform to provide a precise quantification of P. tomentosa DNA and demonstrate the resistance of this assay to inhibitors commonly found in freshwater aquaria.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping