A cross-priming isothermal amplification (CPA) assay was developed for detection of feline herpesvirus type 1 (FHV-1). In this assay, the target fragment of the FHV-1 glycoprotein B gene is amplified rapidly by Bst DNA polymerase at a constant temperature (63 °C, 45 min), using a simple thermostat. The assay had no cross-reactions with four types of feline viruses, and the detection limit was 100 copies/μl. The positive rate of clinical samples from CPA was 100% consistent with qPCR but higher than ordinary PCR, indicating its superiority to ordinary PCR. Visualization was achieved using SYBR Green I dye.
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This work funded was by the National Key Research and Development Program of China (grant number 2016YFD0501002).
Ethical approval and consent to participate
All samples were collected in accordance with the guidelines and regulations of the Animal Care and Use Committee of Jilin Agricultural University, Jilin Province, China. The owners consented to the collection of conjunctival and nasal discharge from the cats.
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Tan, Y., Dong, G., Xu, H. et al. Development of a cross-priming isothermal amplification assay based on the glycoprotein B gene for instant and rapid detection of feline herpesvirus type 1. Arch Virol 165, 743–747 (2020). https://doi.org/10.1007/s00705-020-04526-5