Establishment of a Real-Time PCR Assay for the Detection of Devriesea agamarum in Lizards

(1) Background: ( ) is a potential cause of dermatitis and cheilitis in lizards. The aim of this study was to establish a real-time PCR assay for the detection of . (2) Methods: Primers and probe were selected targeting the 16S rRNA gene, using sequences of 16S rRNA genes of as well as of other bact...

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Bibliographic Details
Published in:Animals (Basel) 2023-02, Vol.13 (5), p.881
Main Authors: Brockmann, Maria, Leineweber, Christoph, Hellebuyck, Tom, Martel, An, Pasmans, Frank, Gentil, Michaela, Müller, Elisabeth, Marschang, Rachel E
Format: Article
Language:English
Subjects:
Annealing
Assaying
Bacteria
Bacterial diseases
bearded dragon
Cell culture
Coefficient of variation
Dermatitis
Devriesea agamarum
Laboratories
lizard
Lizards
Pogona vitticeps
polymerase chain reaction
Process controls
Real time
rRNA 16S
Sheep
Standard deviation
Uromastyx
Uromastyx sp
Veterinary medicine
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Summary:(1) Background: ( ) is a potential cause of dermatitis and cheilitis in lizards. The aim of this study was to establish a real-time PCR assay for the detection of . (2) Methods: Primers and probe were selected targeting the 16S rRNA gene, using sequences of 16S rRNA genes of as well as of other bacterial species derived from GenBank. The PCR assay was tested with 14 positive controls of different cultures as well as with 34 negative controls of various non- bacterial cultures. Additionally, samples of 38 lizards, mostly spp. and spp., submitted to a commercial veterinary laboratory were tested for the presence of using the established protocol. (3) Results: Concentrations of as low as 2 × 10 colonies per mL were detectable using dilutions of bacterial cell culture (corresponding to approximately 200 CFU per PCR). The assay resulted in an intraassay percent of coefficient of variation (CV) of 1.31% and an interassay CV of 1.80%. (4) Conclusions: The presented assay is able to detect in clinical samples, decreasing laboratory turn-around time in comparison to conventional culture-based detection methods.
ISSN:2076-2615
2076-2615
DOI:10.3390/ani13050881