Abstract
As companion animals, felines play an important role in human's family life, and their healthcare has attracted great attention. Viruses such as feline calicivirus (FCV), feline herpesvirus 1 (FHV-1), and feline parvovirus virus (FPV) are the most common pathogens that cause severe infectious disease in baby cats. Thus, preclinical detection and intervention of these three viruses is an effective means to prevent diseases and minimize their danger condition. In this study, a triplex TaqMan quantitative real-time polymerase chain reaction (qRT-PCR) assay was developed to detect these three viruses simultaneously. The detection limit of FPV, FCV, and FHV-1 was 5 × 101 copies/assay, which exhibited higher sensitivity (about 10- to100-fold) than conventional PCR. The coefficients of variation (CVs) of the intra-assay variability were lower than 1.86%, and that of inter-assay variability were lower than 3.19%, indicating the excellent repeatability and reproducibility of the triplex assay. Additionally, the assay showed good specificity. Finally, samples from 48 cats were analyzed using the established assay and commercial kits. As a result, the total positive rates for these viruses were 70.83 or 62.5%, respectively, which demonstrated that the developed qRT-PCR assay was more accurate than the commercial kits and could be used in clinical diagnosis.
Highlights
As companion animals, cats play an important role in human’s life, and their health care has attracted great attention
We find that when the primer concentration is lower than 0.35 μM, the cycle threshold (Ct) value remains at a high level regardless the changes in probe concentration and annealing temperature
The results show that the fluorescence intensities of feline calicivirus (FCV), feline herpesvirus 1 (FHV-1), and feline parvovirus virus (FPV) are at high level when the primer concentration is in the range of 0.5–0.6 μM (Supplementary Figures S1A–C)
Summary
Cats play an important role in human’s life, and their health care has attracted great attention. Infectious disease caused by feline calicivirus (FCV), feline herpesvirus 1 (FHV-1), and feline parvovirus virus (FPV) still pose a threat to cats’ health [1, 2]. FCV is an RNA virus belonging to Vesivirus of Caliciviridae. Studies show that FCV is the most widespread feline virus, especially in multi-cat households and shelters, with overall prevalence ranges from ∼15–31% [3]. The morbidity of FCV can reach to 90% in some colonies [4, 5].
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