ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Infectious Diseases
Volume 12 - 2025 | doi: 10.3389/fvets.2025.1609897
Automated pixel-based quantification of porcine circovirus 2 genome in formalin-fixed, paraffin-embedded tissues using in situ hybridisation
Provisionally accepted
Mònica Sagrera1
Àlex Cobos1Laura Garza-Moreno2Mónica Pérez1Gema García- Buendía1Eva Huerta1Anna Maria Llorens1David Espigares2
Marina Sibila1
Joaquim Segalés1,3*- 1IRTA-CReSA, Centre for Research on Animal Health, Bellaterra, Spain
- 2Ceva Santé Animale, Libourne, Nouvelle-Aquitaine, France
- 3Autonomous University of Barcelona, Barcelona, Spain
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Porcine circovirus 2 (PCV2) detection in lymphoid tissues is paramount for diagnostic purposes. This study developed an automated pixel classifier to quantify PCV2 genome using in situ hybridisation (ISH). Tonsils (TO), tracheobronchial (TBLN), mesenteric (MSLN), and superficial inguinal lymph nodes (ISLN) from 66 experimentally inoculated pigs were analysed with RNAscope ® ISH technology and scored visually (0-3) and digitally (percentage of labelled area). A strong correlation between both methods (ρ=0.96) was confirmed, enabling threshold establishment for digital scores. The TBLN, tissue with the highest PCV2 labelling, was further assessed by PCV2 qPCR, showing high correlation with ISH (ρ=0.85). These findings support the use of digital pathology tools to enhance consistency, reduce subjectivity, and improve efficiency in PCV2 genome quantification in lymphoid tissues.
Keywords: Porcine circovirus 2, In situ hybridisation, qPCR, RNAScope ®, digital pathology
Received: 11 Apr 2025; Accepted: 20 Jul 2025.
Copyright: © 2025 Sagrera, Cobos, Garza-Moreno, Pérez, García- Buendía, Huerta, Llorens, Espigares, Sibila and Segalés. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Joaquim Segalés, Autonomous University of Barcelona, Barcelona, Spain
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