Abstract
Feline chaphamaparvovirus (FeChPV) is a new viral strain detected in Chinese Mainland in recent years. The symptoms mainly include diarrhea and bloody stool in young cats, which can lead to death in severe cases. In this study, a TaqMan-based real-time quantitative PCR (qPCR) with specific primers and TaqMan probes based on the VP1 gene sequence of FeChPV was performed to detect the virus. The established qPCR indicated that there is no cross-reaction of FeChPV with other common feline viruses. The minimum detection limit of the established qPCR method is 3.75 × 10 copies/µL, while conventional PCR is 3.75 × 103 copies/µL. The result that the proposed qPCR protocol was shown to be 100 times more sensitive than conventional PCR. The correlation coefficients exceeded 0.995, and the amplification efficiency was 98%. The difference within and between groups is less than 5%, indicating that the established method has good repeatability. The results of clinical sample detection shown that 16 positive samples were detected from 45 stool samples by the established qPCR method. The conventional PCR method only detected 3 positive samples. In conclusion, the established qPCR method is fast and effective in identifying FeChPV, with higher specificity and sensitivity. It could be used as a diagnostic tool to quantitatively detect the virus content, which is conducive to disease monitoring and epidemiological investigation.
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Data availability
The data that support the findings of this study are available on request from the corresponding author, wangyongcau@126.com, upon reasonable request.
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Acknowledgements
We would like to thank Editage (www.editage.cn) for English language editing. This research is financially funded by Innovation and Entrepreneurship Project of Anhui Agricultural University (national innovation project: 202210364098 and school-level innovation project: X202210364013). Accession Numbers: FeChPV (HF2 isolate, GenBank accession: MT708231.1), Feline parvovirus (FPV, HF1 strain, GenBank accession: MT614366), feline bocavirus 1 (FBov-1, strain AAU01, GenBank accession: MT577646.1), feline bocavirus 3 (FBov-3, GenBank accession: MT633128) and feline herpesvirus (FHV, felo-vaxPCT cat vaccine, Boehringer Ingelheim Group), feline Kobuvirus (FKoV, HFZ-FKV1 strain, GenBank accession: ON219928) and feline calicivirus (FCV, ANHF1 strain, GenBank accession: MT649084).
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SL, XH, and YM were involved in performing the experiments. XL, JW and YC were involved in analyzing data and designing the experiments. YW and SL wrote the manuscript. All the authors have read and approved the final manuscript.
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All experiments were compliant with the ethical standards of Anhui Agricultural University.
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Informed consent was obtained from all the participants included in this study.
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No human studies were carried out by the authors for this article. The animal trials in this study were performed in accordance with the ethical standards of Anhui Agricultural University.
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Li, S., Huo, X., Mu, Y. et al. TaqMan-based real-time polymerase chain reaction for the detection of feline chaphamaparvovirus. 3 Biotech 14, 61 (2024). https://doi.org/10.1007/s13205-024-03917-8
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DOI: https://doi.org/10.1007/s13205-024-03917-8