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

IF 3.9 3区生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Applied Microbiology and Biotechnology Pub Date : 2025-02-14 DOI:10.1007/s00253-024-13394-x

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

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引用次数: 0

Abstract

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/μL, 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.

• One-step duplex TaqMan real-time PCR detection method can detect FCoV and FPLV in clinical samples simultaneously and steadily.

• Almost all the currently known FCoV and FPLV strains can be detected.

• This method has high sensitivity, specificity and generality.

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来源期刊

Applied Microbiology and Biotechnology 工程技术-生物工程与应用微生物

CiteScore

10.00

自引率

4.00%

发文量

535

审稿时长

2 months

期刊介绍： Applied Microbiology and Biotechnology focusses on prokaryotic or eukaryotic cells, relevant enzymes and proteins; applied genetics and molecular biotechnology; genomics and proteomics; applied microbial and cell physiology; environmental biotechnology; process and products and more. The journal welcomes full-length papers and mini-reviews of new and emerging products, processes and technologies.