README file
Data Set Title: “Dataset of dog and cat population tested for Feline panleukopenia virus and Canine parvovirus”
Dataset authors: Veronica Facile (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna); 
Andrea Balboni (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);
Lorenza Urbani (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);
Mara Battilani (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna);

Data Set Contact Persons: Andrea Balboni (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna), email: a.balboni@unibo.it; Veronica Facile (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna), email: veronica.facile2@unibo.it; Mara Battilani (Department of Veterinary Medical Science, Alma Mater Studiorum – University of Bologna), email: mara.battilani@unibo.it 

Data Set License: this data set is distributed under a Creative Commons Attribution 4.0 International (CC BY) license, https://creativecommons.org/licenses/by/4.0/  

Publication Year: 2025

Project Info: 
PRIN2022PNRR_COVDC (Exploring the virome of domestic dogs and cats COVDC (Comprehending virome of domestic carnivores): exploring the viral diversity of domestic carnivores and One Health), Project N° P2022FR49N, PRIN 2022 PNRR funding programme (CUP ID N. J53D23014340001). Funded from the European Union - Next-GenerationEU through the National Recovery and Resilience Plan (NRRP) (MISSION 4 COMPONENT C2, INVESTIMENT N. 1.1), https://www.uniba.it/it/ricerca/dipartimenti/dipmedveterinaria/ricerca/progetti-1/progetti-prin-2022-pnrr/cod-p2022fr49n-exploring-the-virome-of-domestic-dogs-and-cats-covdc-comprehending-virome-of-domestic-carnivores-exploring-the-viral-diversity-of-domestic-carnivores-and-one-health-implications 
INF-ACT (One Health Basic and Translational Research Actions addressing Unmet Needs on Emerging Infectious Diseases), funded by European Union, NextGenerationEU-MUR PNRR Partnership initiative on Emerging Infectious Diseases Programme (PNRR, Missione 4 Componente 2 - Investimento 1.3). Project number PE00000007, CUP ID B13C22002440006; https://www.inf-act.it/
Data set Contents
The data set consists of:
•	1 tabular qualitative file in .xlsx format
PRIN2022PNRR_P2022FR49N-COVDC_DatasetHRMParvovirus_2025.xlsx
•	1 readme file in .rtf format
PRIN2022PNRR_P2022FR49N-COVDC_DatasetHRMParvovirus_2025_README.rtf

Data set Documentation
Abstract
The dataset contains signalment, clinical and diagnostic data of a population of 33 dogs and cats tested for the presence of feline panleukopenia virus and canine parvovirus DNA at the Veterinary Teaching Hospital of the Department of Veterinary Medical Science of the University of Bologna.

Content of the files 
file PRIN2022PNRR_P2022FR49N-COVDC_DatasetHRMParvovirus_2025.xlsx contains signalment, clinical and diagnostic data on 33 dogs and cats tested for the presence of feline panleukopenia virus and canine parvovirus DNA at the Veterinary Teaching Hospital of the Department of Veterinary Medical Science of the University of Bologna. Specifically, information about the breed, the origin, the age and year of birth, the sex, the year of sampling, the vaccination status, the presence of clinical signs correlated with parvovirus infection and the qPCR, genotyping (columns V, W and Y) and sequencing (column T) results are indicated for each tested animal. The first qPCR-HRM assay (qPCR-HRM-1) was carried out with the intent of differentiating FPV, original CPV-2 and CPV-2 antigenic variants (2a, 2b and 2c). The second qPCR-HRM assay (qPCR-HRM-2), was carried out only on CPV-positive samples to differentiate Asian-like CPV strains from other CPVs. 

Methodologies 
The two qPCR assays used to perform HRM analysis were developed and validated by the authors. Reactions were performed using the QuantStudio™ 3 Real-Time PCR System (Thermo Fisher Scientific, Applied Biosystems, Waltham, MA, USA) with a total reaction volume of 50 μl per tube. Each tube contained 25 μl Applied Biosystems MeltDoctor reagents (Thermo Fisher Scientific, Life Technologies, Waltham, MA, USA), 3 μl of forward and reverse primers, respectively, with a concentration of 5 µM each, 2.5 μl of DNA template, and 16.5 μl of water for molecular biology. Each sample was tested in triplicates. The cycling conditions included a denaturation stage at 95 °C for 10 min, followed by the PCR stage of 40 cycles at 95 °C for 15 s and 60 °C for 1 min. The determination of melting curve was performed as follows: 95 °C for 10 s, 60 °C for 1 min to 95 °C for 15 s at a rate of 0.025 °C/s. HRM analysis was conducted using the Applied Biosystems High Resolution Melt Software v3.1 (Thermo Fisher Scientific, Waltham, MA, USA). The software automatically assigned samples to variants based on melting curve similarity with positive control.
All viruses identified and typed with the developed assays were subsequently sequenced for confirmation and genetic characterization. For this purpose, a previously described end-point PCR (Balboni et al., 2021)1, amplifying the entire VP2 gene, was carried out using the Phusion Hot Start II High-Fidelity DNA Polymerase Kit (Thermo Fisher Scientific, Life Technologies, Waltham, MA, USA) according to the manufacturer’s instructions. Amplicons of the expected size obtained with the PCR assay carried out in this study were sequenced by Sanger method using both forward and reverse primers.
 Balboni A, Niculae M, Di Vito S, Urbani L, Terrusi A, Muresan C, Battilani M. The detection of canine parvovirus type 2c of Asian origin in dogs in Romania evidenced its progressive worldwide diffusion. BMC Vet Res. 2021 Jun 5;17(1):206. doi: 10.1186/s12917-021-02918-6.