The establishment and application of a dual Nano-PCR detection method for feline calicivirus and feline herpesvirus type I

Manping Yan; Jinyuan Shang; Xiaohao Zhang; Shun Wu; Chunxia Wang; Zhenjun Wang; Guoliang Luo; Li Yi; Xiaofeng Shan; Yuening Cheng; Erkai Feng

doi:10.3389/fmicb.2023.1285268

The establishment and application of a dual Nano-PCR detection method for feline calicivirus and feline herpesvirus type I

Front Microbiol. 2023 Nov 16:14:1285268. doi: 10.3389/fmicb.2023.1285268. eCollection 2023.

Authors

Manping Yan^{1

2}, Jinyuan Shang¹, Xiaohao Zhang³, Shun Wu¹, Chunxia Wang¹, Zhenjun Wang¹, Guoliang Luo¹, Li Yi¹, Xiaofeng Shan², Yuening Cheng¹, Erkai Feng¹

Affiliations

¹ Key Laboratory of Economic Animal Diseases, Ministry of Agriculture, Institute of Special Animal and Plant Sciences, Chinese Academy of Agricultural Sciences, Changchun, China.
² College of Animal Science and Technology, Jilin Agricultural University, Changchun, China.
³ Department of Cardiology, The Second Hospital of Jilin University, Changchun, China.

Abstract

Feline calicivirus (FCV) and Feline herpesvirus type I (FHV-I) are the main pathogens causing upper respiratory tract infections in cats, and some wild animals. These two viruses always coinfection and cause serious harm to pet industry and wild animals protection. Established a rapid and accurate differential diagnosis method is crucial for prevention and control of disease, however, the current main detection method for these two viruses, either is low sensitivity (immunochromatographic strip), or is time-consuming and cannot differential diagnosis (conventional single PCR). Nanoparticle-assisted polymerase chain reaction (Nano-PCR) is a recently developed technique for rapid detection method of virus and bacteria. In this study, we described a dual Nano-PCR assay through combining the nanotechnology and PCR technology, which for the clinical simultaneous detection of FCV and FHV-I and differential diagnosis of upper respiratory tract infections in cats or other animals. Under optimized conditions, the optimal annealing temperature for dual Nano-PCR was 51.5°C, and specificity test results showed it had no cross reactivity to related virus, such as feline panleukopenia virus (FPV), feline Infectious peritonitis virus (FIPV) and rabies virus (RABV). Furthermore, the detection limit of dual Nano-PCR for FCV and FHV-I both were 1 × 10^-8 ng/μL, convert to number of copies of virus DNA was 6.22 × 10³copies/μL (FCV) and 2.81 × 10³copies/μL (FHV-I), respectively. The dual Nano-PCR detected result of 52 cat clinical samples, including ocular, nasal and faecal swabs, and (3 FCV-positive samples), was consistent with ordinary PCR and the clinical detection results. The dual Nano-PCR method established in this study with strong specificity and high sensitivity can be used for virus nucleic acid (FCV and FHV-I) detection of clinical samples of feline upper respiratory tract infections feline calicivirus and feline herpesvirus while providing support for the early diagnosis of cats that infected by FCV and FHV-I.

Keywords: Nano-PCR; detection diagnostic technology; feline calicivirus; feline herpesvirus type I; infectious disease.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by the Science and Technology Development Plan of Jilin Province (No. 20210202099NC), Innovation Project of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-2021-ISAPS) and Basic Construction Funds within the budget of Jilin Province (No. 2022C042-9).