Visible and rapid detection of feline chaphamaparvovirus using multienzyme isothermal rapid amplification and lateral flow dipstick assay.

IF 4.8 2区医学 Q2 IMMUNOLOGY Frontiers in Cellular and Infection Microbiology Pub Date : 2025-01-22 eCollection Date: 2025-01-01 DOI:10.3389/fcimb.2025.1490948

Jun Ji, Xinhao Mu, Shunshun Pan, Xin Xu, Shiyuan Zhang, Honghui Huang, Ying Li, Yingzuo Bi, Lunguang Yao

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Abstract

Feline chaphamaparvovirus (FeChPV) is a novel parvovirus previously reported in Canadian cats and Chinese dogs with diarrhea in 2019 and 2020, respectively. Herein, we aimed to establish a simple detection method for FeChPV in field clinics. The primers and probes for the multienzyme isothermal rapid amplification and lateral flow dipstick (MIRA-LFD) assay were designed to target the conserved regions of the FeChPV genome and determine the optimal reaction temperature and time. Without relying on precision instruments, FeChPV detection using the MIRA-LFD assay was completed within 20 min at 37°C, without any cross-reaction with other reference viruses. The newly established MIRA-LFD assay had a detection limit of 32.3 copies/μL, which was 10-fold lower than that of the nested polymerase chain reaction (PCR) assay. Furthermore, the MIRA-LFD assay detected 29 FeChPV-positive samples among 417 cats with diarrhea, providing a slightly higher positivity rate than the nested PCR assay. These results indicate that the newly developed MIRA-LFD assay for FeChPV detection is an efficient, economical, reliable, and simple method that can help in the early prevention and control of FeChPV infection.

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采用多酶等温快速扩增和侧流试纸法对猫chaphamaparv病毒进行可见快速检测。

猫chaphamparvovirus （FeChPV）是一种新型细小病毒，此前分别于2019年和2020年在加拿大猫和中国狗身上报道过腹泻。在此，我们旨在建立一种简便的现场临床FeChPV检测方法。多酶等温快速扩增和侧流量尺（MIRA-LFD）检测的引物和探针旨在针对FeChPV基因组的保守区域，并确定最佳反应温度和时间。在不依赖精密仪器的情况下，使用MIRA-LFD法检测FeChPV在37°C下20分钟内完成，与其他参考病毒无交叉反应。新建立的MIRA-LFD法的检出限为32.3 copies/μL，比巢式PCR法的检出限低10倍。此外，MIRA-LFD法在417只腹泻猫中检测到29份fechpv阳性样本，阳性率略高于巢式PCR法。这些结果表明，新开发的MIRA-LFD检测FeChPV是一种高效、经济、可靠、简单的方法，可以帮助FeChPV感染的早期预防和控制。

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来源期刊

Frontiers in Cellular and Infection Microbiology IMMUNOLOGY-MICROBIOLOGY

CiteScore

7.90

自引率

7.00%

发文量

1817

审稿时长

14 weeks

期刊介绍： Frontiers in Cellular and Infection Microbiology is a leading specialty journal, publishing rigorously peer-reviewed research across all pathogenic microorganisms and their interaction with their hosts. Chief Editor Yousef Abu Kwaik, University of Louisville is supported by an outstanding Editorial Board of international experts. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide. Frontiers in Cellular and Infection Microbiology includes research on bacteria, fungi, parasites, viruses, endosymbionts, prions and all microbial pathogens as well as the microbiota and its effect on health and disease in various hosts. The research approaches include molecular microbiology, cellular microbiology, gene regulation, proteomics, signal transduction, pathogenic evolution, genomics, structural biology, and virulence factors as well as model hosts. Areas of research to counteract infectious agents by the host include the host innate and adaptive immune responses as well as metabolic restrictions to various pathogenic microorganisms, vaccine design and development against various pathogenic microorganisms, and the mechanisms of antibiotic resistance and its countermeasures.