ORIGINAL RESEARCH article
Front. Vet. Sci.
Sec. Veterinary Clinical, Anatomical, and Comparative Pathology
This article is part of the Research TopicInnovations in Veterinary Diagnostics for Enhancing Animal Health and WelfareView all 17 articles
Automated highly multiplex detection system for respiratory pathogens in canine
Provisionally accepted
- 1EMERGING VIRAL DIAGNOSTICS (HK) LIMITED, Hong Kong, Hong Kong, SAR China
- 2Hong Kong Baptist University, Hong Kong, Hong Kong, SAR China
- 3The Hong Kong Polytechnic University, Hong Kong, Hong Kong, SAR China
Select one of your emails
You have multiple emails registered with Frontiers:
Notify me on publication
Please enter your email address:
If you already have an account, please login
You don't have a Frontiers account ? You can register here
Canine Infectious Respiratory Diseases (CIRD) are prevalent causes of respiratory illnesses in dogs. Clinical signs are non-specific, including coughing, rhinorrhea, and fever, making it challenging for veterinarians, especially at onset of symptoms, to identify the causative pathogens based on clinical presentation alone. On the other hand, early and accurate diagnosis is crucial for preventing progression to severe complications such as pneumonia and widespread outbreaks. The ability to differentiate between viral and bacterial etiologies can guide appropriate treatment and medication directions, such as avoiding mis-use of antibiotics. Therefore, this work aims to develop a novel multiple molecular assay suitable for an automated detection system using a nested PCR method. The assay covers 14 common canine respiratory pathogens using 15 gene targets, including Canine Influenza Virus (H3N2, H3N8, H1N1), Canine Distemper Virus, Canine Parainfluenza Virus, Canine Herpesvirus, Pseudorabies Virus, Rabies Virus, Canine Adenovirus (Types 1 and 2), Canine Coronavirus, Mycoplasma canis, Bordetella bronchiseptica, and Streptococcus equi subsp. zooepidemicus. Their primers and probes in the assay were first developed according to the nested PCR protocol and designed parameters required in the automated system. This developed assay has then been rigorously validated. It has demonstrated high analytical sensitivity and specificity. Results obtained from the automated system are comparable to those from conventional laboratory procedures. The assay has showed possibility in detecting rapidly multiple pathogens in canine and its utility can potentially be extended beyond companion animals to other mammalian species as well. Its application can enhance infection surveillance in animal populations and potentially mitigate zoonotic transmission risks.
Keywords: Multiplex nested PCR, CIRD, Canine respiratory pathogens, Automation, Point - of - care diagnostics
Received: 10 Oct 2025; Accepted: 24 Nov 2025.
Copyright: © 2025 Wong, Lin, Tsang, Lau and Lau. 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: Lok-Ting Lau, terencelau@hkbu.edu.hk
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.