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犬圆环病毒芯片数字PCR检测方法的建立

Establishment of a chip digital PCR detection method for canine circovirus.

作者信息

Lei Xiaoxiao, Lv Qiao, Qin Yan, Chen Wei, Hu Yanqing, Zhao Chenchen, Zhang Xinyu, Huang Haixin, Li Yuying, Lu Jingyi, Lan Tian, Sun Wenchao, Zheng Min

机构信息

Wenzhou Key Laboratory for Virology and Immunology, Institute of Virology, Wenzhou University, Wenzhou, 325035, China.

Guangxi Centre for Animal Disease Control and Prevention, Nanning, 530001, China.

出版信息

Heliyon. 2024 May 9;10(9):e30859. doi: 10.1016/j.heliyon.2024.e30859. eCollection 2024 May 15.

Abstract

Canine circovirus (CanineCV), which is a new mammalian circovirus first reported in the United States in 2012, mainly causes diarrhea and vomiting in dogs. As CanineCV evolves and new subtypes emerge, there is an urgent need for new detection technologies to improve the sensitivity and detection rates of viruses in complex scenarios. A chip digital PCR(cdPCR) assay was established for the detection of CanineCV in this study. The results showed good reproducibility, specificity and a linear relationship; the minimum detection limit of CanineCV by cdPCR was 6.62 copies/μL, which is 10 times more sensitive than quantitative real-time PCR (qPCR). The qPCR-positive detection rate was 1 %, while CanineCV cdPCR (2.1 %) exhibited a greater positive detection rate. Fifteen complete genomes were sequenced and subdivided into CanineCV-1 and CanineCV-3. In conclusion, we developed a rapid, reliable, and specific cdPCR method for screening and monitoring canine CV.

摘要

犬圆环病毒(CanineCV)是2012年首次在美国报道的一种新型哺乳动物圆环病毒,主要引起犬类腹泻和呕吐。随着CanineCV的进化和新亚型的出现,迫切需要新的检测技术来提高复杂情况下病毒的敏感性和检测率。本研究建立了一种用于检测CanineCV的芯片数字PCR(cdPCR)检测方法。结果显示出良好的重复性、特异性和线性关系;cdPCR检测CanineCV的最低检测限为6.62拷贝/μL,比定量实时PCR(qPCR)灵敏10倍。qPCR阳性检测率为1%,而CanineCV cdPCR(2.1%)显示出更高的阳性检测率。对15个完整基因组进行了测序,并细分为CanineCV-1和CanineCV-3。总之,我们开发了一种快速、可靠且特异的cdPCR方法用于犬圆环病毒的筛查和监测。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd4d/11107241/9ed1fdfe3776/gr1.jpg

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