Establishment of one-step duplex TaqMan real-time PCR for detection of feline coronavirus and panleukopenia virus

文献类型： 外文期刊

第一作者： Liu, Zhe

作者： Liu, Zhe;Jiang, Qian;Yang, Yupeng;Qi, Ruibin;Gu, Haorong;Feng, Kexin;Jia, Honglin;Kang, Hongtao;Liu, Jiasen;Chen, Mengru

作者机构：

关键词： Duplex TaqMan qPCR; FCoV; FPLV; Clinical detection

期刊名称：APPLIED MICROBIOLOGY AND BIOTECHNOLOGY （ 影响因子：4.3； 五年影响因子：5.1 ）

ISSN： 0175-7598

年卷期： 2025 年 109 卷 1 期

页码：

收录情况： SCI

摘要： A comparative genomic analysis of feline coronavirus (FCoV) and feline panleukopenia virus (FPLV) was performed. Based on the conserved regions of the two viruses, specific probes and real-time PCR (qPCR) primers were designed, and a duplex TaqMan qPCR-based assay was established for detecting FCoV and FPLV. The results showed high analytical specificity, and no cross-response with other feline viruses was observed. This method is highly versatile and can be used to detect all FCoV strains stored in laboratories and recombinant plasmids constructed according to the sequences of blank FCoV strains in laboratories. The analytical sensitivity of this method in detecting FCoV and FPLV was as low as 50 copies/mu L, which is approximately 20-fold greater than that of conventional PCR. The coefficients of variation (CVs) for the intra- and interbatch coefficients of variation were less than 2%. After 75 clinical samples were tested, the percentage of FCoV- and FPLV-positive samples was 5.34% greater than that of conventional PCR methods, a finding robustly supported by sequencing identification. As validated by clinical samples, the method was sensitive, specific, general, and reproducible and holds great potential for the rapid identification and diagnosis of FCoV and FPLV infections, as well as for epidemiological investigations.

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