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Research Article | 03 Oct 2026

Quantitative polymerase chain reaction detection of hemotropic and vector-borne pathogens in dogs and cats using blood and pooled blood–urine matrixes: A large-scale retrospective study in Colombia

Juan Fernando Manrique-Hincapie1, William Armando Tapie1, Juan Esteban Lamprea Rodríguez2, Leidy Alejandra Giraldo Martínez2, and César Orlando Muñoz Cadavid3 Show more
VETERINARY WORLD | Article No. 1 | pg no. 4259-4273 | Vol. 19, Issue 10 | DOI: 10.14202/vetworld.2026.4259-4273
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ABSTRACT

                      
Background and Aim: Hemotropic and vector-borne pathogens are important causes of disease in dogs and cats, particularly in tropical regions. Whole blood is conventionally used for molecular diagnosis; however, fluctuating pathogen loads may reduce detection. Urine could provide complementary pathogen-derived DNA, but evidence supporting pooled blood–urine (BU) matrixes remains limited. This study compared whole blood with a pooled BU matrix for quantitative polymerase chain reaction (qPCR) detection of hemotropic and vector-borne pathogens in dogs and cats in Colombia.
Materials and Methods: This retrospective study included 12,491 unique animals (8,169 dogs and 4,322 cats) tested from January to December 2025. The dataset comprised 11,461 blood and 1,030 pooled BU samples, with the latter standardized at 60% blood and 40% urine. Multipathogen qPCR panels were disaggregated into 100,159 pathogen-specific analytical records. Bacterial agents were detected using assays targeting the 16S rRNA gene, whereas protozoan and parasitic agents were assessed using the 18S rRNA gene. Detection probabilities were analyzed separately for dogs and cats using binomial generalized linear models, with matrix type, pathogen, and geographic origin as fixed effects.
Results: Matrix type did not significantly affect detection probability in dogs (odds ratio [OR] = 1.014, 95% confidence interval [CI] = 0.909–1.128; p > 0.05) or cats (OR = 0.883, 95% CI = 0.689–1.115; p > 0.05). In dogs, Anaplasma spp. positivity rate was 27.32% in blood and 25.40% in pooled BU samples. In cats, Mycoplasma spp. positivity rate was 15.28% and 11.87%, respectively. Geographic origin significantly influenced detection probability (p ≤ 0.05); compared with Bogotá, Santander showed higher odds of positivity in dogs (OR = 1.888) and cats (OR = 1.926).
Conclusion: At the population level, pooled BU samples showed no significant difference from whole blood in qPCR detection probability. These findings support further evaluation of pooled matrixes as a complementary sampling strategy. However, the retrospective, unpaired design and unequal matrix sample sizes preclude conclusions regarding individual level diagnostic equivalence; prospective paired-sample studies are required.
                      
Keywords: blood–urine matrix, cats, Colombia, dogs, hemotropic pathogens, molecular diagnosis, qPCR, vector-borne pathogens.