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Animals
Special Issues
Advances in Molecular Diagnostics in Veterinary Sciences
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Advances in Molecular Diagnostics in Veterinary Sciences

A special issue of Animals (ISSN 2076-2615). This special issue belongs to the section "Veterinary Clinical Studies".

Deadline for manuscript submissions: 30 June 2026 | Viewed by 2601

Special Issue Editors

Dr. Ailam Lim

E-Mail Website
Guest Editor
Wisconsin Veterinary Diagnostic Laboratory, Department of Pathobiological Sciences, University of Wisconsin-Madison, Madison, WI, USA
Interests: molecular; virology; diagnostics; PCR; next-generation sequencing; method development
Dr. Lifang Yan

E-Mail Website
Guest Editor
Mississippi Veterinary Research & Diagnostic Lab, Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University, Pearl, MS, USA
Interests: molecular; virology; diagnostics; PCR; next-generation sequencing; method development

Special Issue Information

Dear Colleagues,

Molecular techniques have become a cornerstone of rapid diagnostics in veterinary sciences. Advances in molecular diagnostics—from cloning and polymerase chain reaction (PCR) to Sanger sequencing, Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR), and next-generation sequencing (NGS)—have equipped researchers and clinicians with powerful tools for detecting infectious diseases, including viral, bacterial, and protozoal infections, as well as neoplastic, metabolic-degenerative, and hereditary disorders. Animals—from pets and livestock to wildlife—play an integral role in ecosystems and are deeply connected to human life. Many animal diseases are zoonotic, posing significant threats to public health at the animal–human interface. The threat of transboundary diseases is at an all-time high, with the global spread of high-consequence diseases leading to significant economic losses. The application of highly sensitive and rapid diagnostic technologies ensures the timely detection of diseases, which is essential for effective management and intervention strategies to protect both animal and public health. This Special Issue aims to highlight the latest advancements and applications of molecular diagnostics in veterinary sciences. We welcome you to submit your work, including original research articles and review papers that offer new perspectives on molecular diagnostics for infectious diseases in veterinary medicine.

We look forward to receiving your contributions.

Dr. Ailam Lim
Dr. Lifang Yan
Guest Editors

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 250 words) can be sent to the Editorial Office for assessment.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Animals is an international peer-reviewed open access semimonthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2400 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • polymerase chain reaction (PCR)
  • next-generation sequencing (NGS)
  • molecular diagnostics
  • infectious diseases
  • transboundary diseases
  • CRISPR

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Further information on MDPI's Special Issue policies can be found here.

Published Papers (2 papers)

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Research

17 pages, 5992 KB  
Article
Development and Evaluation of Quadruplex Droplet Digital PCR Method to Multiplex Detection of Different Respiratory Pathogens of Chickens
by Yingli Mu, Xuejing Wang, Tongchao Dong, Xinran Bao, Qianqian Xu, Tianxiang Lan, Juxiang Liu and Ligong Chen
Animals 2026, 16(1), 139; https://doi.org/10.3390/ani16010139 - 3 Jan 2026
Viewed by 859
Abstract
Chicken respiratory diseases represent multifactorial conditions resulting from viral, bacterial, mycoplasmal pathogens, and environmental factors, causing significant economic losses within the poultry industry. A specific respiratory disease characterized by breathing difficulties and bronchial occlusion due to caseous exudates is termed chicken bronchial obstruction. [...] Read more.
Chicken respiratory diseases represent multifactorial conditions resulting from viral, bacterial, mycoplasmal pathogens, and environmental factors, causing significant economic losses within the poultry industry. A specific respiratory disease characterized by breathing difficulties and bronchial occlusion due to caseous exudates is termed chicken bronchial obstruction. However, the absence of rapid, precise, and highly sensitive diagnostic methods for differentiation of primary respiratory disease pathogens or opportunistic pathogens, including avian influenza virus (AIV), infectious bronchitis virus (IBV), Pseudomonas aeruginosa (P. aeruginosa), and Escherichia coli (E. coli), constitutes a substantial challenge. This study developed a quadruplex droplet digital polymerase chain reaction (ddPCR) method that targeted the HA gene of H9 subtype AIV, the M gene of IBV, the Pal gene of P. aeruginosa, and the UidA gene of E. coli. Following the optimization of annealing temperature, sensitivity, and repeatability, the minimum detectable concentrations were determined as 3.02 copies/μL for the HA gene of H9 subtype AIV, 3.08 copies/μL for the M gene of IBV, 3.19 copies/μL for the Pal gene of P. aeruginosa, 3.39 copies/μL for the UidA gene of E. coli. No cross-reactivity was observed with Newcastle disease virus (NDV), H5 subtype AIV, H7 subtype AIV, fowl adenovirus serotype 4 (FAdV-4), infectious laryngotracheitis virus (ILTV), Avibacterium paragallinarum, Streptococcus, Salmonella, Pasteurella multocida, and Staphylococcus aureus. The method demonstrated excellent repeatability, with a coefficient of variation (CV) below 9%. The 185 clinical samples collected in Hebei Province China are tested by both quadruplex ddPCR and quadruplex qPCR method and the results compared. The sensitivity of the quadruplex ddPCR method (57.30%; 106/185) slightly exceeded that of the quadruplex qPCR method (49.73%; 92/185). Pathogens or opportunistic pathogens positive rates obtained via the quadruplex ddPCR were 40.00% for H9 subtype AIV, 33.51% for IBV, 24.32% for P. aeruginosa, and 27.57% for E. coli. In comparison, the positive rates of H9 subtypes AIV, IBV, P. aeruginosa, and E. coli from the quadruplex qPCR were 36.22%, 30.81%, 21.62%, and 24.32%, respectively. The coincidence rates between the two methods were 96.22% for H9 AIV, 97.30% for IBV, 97.30% for P. aeruginosa, and 96.76% for E. coli. These results demonstrated that the quadruplex ddPCR method represented a highly sensitive, specific, and rapid technique for identifying H9 subtype AIV, IBV, P. aeruginosa, and E. coli. Full article
(This article belongs to the Special Issue Advances in Molecular Diagnostics in Veterinary Sciences)
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17 pages, 2003 KB  
Article
Optimizing Nucleic Acid Extraction from Extended Bovine Semen for Endemic and High-Consequence Pathogens
by Amanda Zimmerman, Anne Vandenburg-Carroll, Douglas G. Marthaler and Ailam Lim
Animals 2025, 15(23), 3411; https://doi.org/10.3390/ani15233411 - 26 Nov 2025
Cited by 1 | Viewed by 683
Abstract
Accurate pathogen detection in bovine semen is crucial for animal health surveillance and international trade. Semen presents unique challenges due to the presence of PCR inhibitors from seminal plasma and extender components, reducing nucleic acid extraction efficiency and sensitivity. The two National Animal [...] Read more.
Accurate pathogen detection in bovine semen is crucial for animal health surveillance and international trade. Semen presents unique challenges due to the presence of PCR inhibitors from seminal plasma and extender components, reducing nucleic acid extraction efficiency and sensitivity. The two National Animal Health Laboratory Network-approved extraction platforms (MagMAX CORE and IndiMag Pathogen Kits) were evaluated using 88 negative extended semen samples at 200 µL input volume, reduced input volumes, and pretreatment strategies with two influenza A virus (IAV) PCR assays, containing different exogenous internal controls (ICs) to assess PCR inhibition. The ICs yielded overall passing rates from 31.8% to 100.0% and varied greatly based on the extender formulation and extraction protocol. Validation continued with naturally infected semen containing Mycoplasma bovis, bovine viral diarrhea virus, bovine herpesvirus-1, and the limit of detection using Mycoplasma bovis. The IndiMag Pathogen 100-na was then selected for evaluation of diagnostic sensitivity and specificity, reproducibility, and detection limits with IAV-spiked samples, using the two IAV PCR assays and their ICs. Selected archived semen samples used in this study were screened and were negative for IAV by both PCR assays. These findings underscore the importance of tailored extraction methods in overcoming semen-associated inhibition and facilitating reliable pathogen surveillance in bovine germplasm. Full article
(This article belongs to the Special Issue Advances in Molecular Diagnostics in Veterinary Sciences)
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