Evaluation of Ante-Mortem Sampling Matrices for qPCR Detection of Canine Distemper Virus
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Abstract
Canine distemper virus (CDV) is a multi-host morbillivirus of domestic dogs and wild carnivores, and early molecular detection is important for treatment, isolation, and wildlife surveillance. However, the most informative ante-mortem specimen type for RT-qPCR screening remains context dependent. This pilot study compared CDV RNA detection and viral load across paired sample matrices from nine dogs and screened archived blood/serum samples from four tigers and one clouded leopard. CDV RNA was extracted using a commercial viral nucleic-acid kit and amplified using a one-step RT-qPCR assay. Five of nine dogs were CDV-positive. Among positive dogs, conjunctival swabs detected CDV RNA in 5/5 animals (100%), followed by nasal swabs (4/5), rectal swabs (3/5), whole blood (2/5), and urine (1/5). Viral load ranged from 9.55 to 213,796.21 RNA copies/μL. Exact Cochran’s Q and Friedman tests did not reach statistical significance, but Kendall’s W suggested a moderate sample-type effect. These results support conjunctival swabs as the most consistent specimen in this dataset, with nasal and rectal swabs as useful complementary matrices.
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This Zenodo record is a permanently preserved version of a PREreview. You can view the complete PREreview at https://prereview.org/reviews/22958187.
The baseline strategy of evaluating clinical sample types to optimize qPCR diagnostic sensitivity grids for Canine Distemper Virus is highly relevant to contemporary mole ular virology. Independent validation of different antemortem matrices is historically neglected, leading to false negatives during acute viral shielding windows.
While the comparative approach across urine, conjunctival, and whole blood fractions is structurally useful, the underlying experimental design requires tighter calibration boundaries regarding extraction variance. The authors must fully report the specific standard deviations and intra-assay coefficients of variation observed across high-density Ct values. …
This Zenodo record is a permanently preserved version of a PREreview. You can view the complete PREreview at https://prereview.org/reviews/22958187.
The baseline strategy of evaluating clinical sample types to optimize qPCR diagnostic sensitivity grids for Canine Distemper Virus is highly relevant to contemporary mole ular virology. Independent validation of different antemortem matrices is historically neglected, leading to false negatives during acute viral shielding windows.
While the comparative approach across urine, conjunctival, and whole blood fractions is structurally useful, the underlying experimental design requires tighter calibration boundaries regarding extraction variance. The authors must fully report the specific standard deviations and intra-assay coefficients of variation observed across high-density Ct values. Bypassing these numerical data limits introduces significant noise into target copy number estimations.
Additionally, tracking target sequence mutations that dynamically alter primer-bining kinetics under selective host pressures is essential to prove long-term assay validation. This technical adjustment establishes an objective benchmark for diagnostic networks mapping cross-host morbillivirus transmission tracks without dry-lab limitations.
Competing interests
The author declares that they have no competing interests.
Use of Artificial Intelligence (AI)
The author declares that they did not use generative AI to come up with new ideas for their review.
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