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Development and Validation of a Droplet Digital PCR Assay for Detection of Feline Herpesvirus Type-1

文献类型： 外文期刊

作者： Yaxi Zhou;Danni Wu;Mengle Tang;Zihan Ye;Erkai Feng;Haili Zhang;Guoliang Luo;Zhenjun Wang;Chunxia Wang;Lina Liu;Yuening Cheng

作者机构：

关键词： droplet digital PCR;feline herpesvirus type-1;gD gene;real-time quantitative PCR

期刊名称： Veterinary Sciences

ISSN： 2306-7381

年卷期： 2025 年 12 卷 11 期

页码：

收录情况： SCIE（2025版）

摘要： Feline herpesvirus type-1 (FHV-1), a double-stranded DNA virus, which is a highly infectious upper respiratory tract infection of felids, particularly in kittens. Droplet digital PCR (ddPCR) provides an absolute quantification method with high sensitivity and accuracy. This study aimed to develop a highly sensitive and accurate ddPCR assay for the detection of FHV-1. We designed primers and a probe targeting the FHV-1 glycoprotein D (gD) gene and evaluated the assay’s limit of detection (LOD), sensitivity, repeatability, and specificity in comparison to quantitative real-time PCR (qPCR). The developed ddPCR assay demonstrated a strong linear dynamic range (R2 ≥ 0.99) and an exceptionally low LOD of 0.18 copies/μL, which was significantly more sensitive than the method qPCR (LOD ~10 copies/μL). Additionally, the assay exhibited high specificity with no cross-reactivity against other common feline pathogens (feline calicivirus, FCV; feline panleukopenia virus, FPV; feline infectious peritonitis virus, FIPV; Bordetella bronchiseptica and Chlamydia felis) and displayed outstanding repeatability (inter-run CV < 1.35). When applied to 118 clinical samples, the ddPCR assay achieved a significantly higher positive detection rate (27.4%) compared to qPCR (14.8%). In conclusion, we have successfully established a reliable ddPCR assay for the absolute quantification of FHV-1, providing a superior tool for laboratory diagnosis and research.

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