Peer-reviewed veterinary case report
Development of a quantitative PCR for the detection of Rangelia vitalii.
- Journal:
- Veterinary parasitology
- Year:
- 2016
- Authors:
- Paim, Francine Chimelo et al.
- Affiliation:
- Laboratory of Veterinary Clinical Analysis · Brazil
- Species:
- dog
Plain-English summary
This study focused on creating a reliable test to find and measure a specific part of the Rangelia vitalii parasite in dog blood. The researchers developed a special method called SYBR Green qPCR, which was able to detect very small amounts of the parasite's DNA without confusing it with other similar organisms. They tested blood samples from 265 dogs in Brazil and found that 24 of them, or about 9%, were positive for R. vitalii. Half of these positive results were confirmed through further testing. Overall, the new test is effective and can help veterinarians diagnose R. vitalii infections in dogs.
Abstract
The aim of this study was to develop and validate a SYBR Green qPCR assay to detect and quantify a fragment of the 18S rRNA gene of Rangelia vitalii in canine blood. Repeatability of the qPCR was determined by the intra- and inter-assay variations. The qPCR showed efficiency of E=101.30 (r(2)=0.996), detecting as few as one copy of plasmid containing the target DNA. Specificity of the assay was performed using DNA samples of Babesia canis, B. gibsoni, Ehrlichia canis, E. ewingii and Leishmania sp. No cross-reactivity was observed. Field samples consisting of blood from 265 dogs from Porto Alegre, Brazil were also tested. A total of 24 (9.05%) samples were positive for R. vitalii. Amplicons of 50% of positive samples were confirmed to be R. vitalii by Sanger sequencing. The positive samples had an average of 3.5×10(5) organisms/mL of blood (range: 1.27×10(3)-1.88×10(6)) based on the plasmid-generated standard curve. In conclusion, the SYBR Green qPCR assay developed herein is sensitive and specific and can be used as a diagnostic tool for detection and quantification of R. vitalii in canine blood samples.
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Search related cases →Original publication: https://pubmed.ncbi.nlm.nih.gov/26827871/