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开发一种用于检测猪圆环病毒 4 的微滴式数字 PCR 方法。

Development of a droplet digital PCR method for detection of porcine circovirus 4.

机构信息

Henan Provincal Engineering and Technology Center of Health Products for Livestock and Poultry, School of Life Science and Agricultural Engineering, Nanyang Normal University, Nanyang, 473061, Henan, China.

College of Veterinary Medicine, Northwest A&F University, Yangling, 712100, Shaanxi, China.

出版信息

BMC Vet Res. 2023 Aug 22;19(1):129. doi: 10.1186/s12917-023-03690-5.

DOI:10.1186/s12917-023-03690-5
PMID:37608311
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10464377/
Abstract

BACKGROUND: Porcine circovirus 4 (PCV4), a newly emerging virus that was first discovered in 2019, may pose a potential threat to the pig industry. Droplet digital PCR (ddPCR) is an absolute quantitative method that has high sensitivity and accuracy. In this study, we developed a novel ddPCR assay to detect PCV4. Furthermore, we evaluated the detection limit, sensitivity, specificity and reproducibility of the ddPCR and TaqMan real-time quantitative PCR (qPCR) and tested 160 clinical samples to compare the detection rate of the two methods. RESULTS: The detection limit for ddPCR was 0.54 copies/µL, 10.6 times greater sensitivity than qPCR. Both ddPCR and qPCR assays exhibited good linearity and repeatability, and the established ddPCR method was highly specific for PCV4. The results of clinical sample testing showed that the positivity rate of ddPCR (5.6%) was higher than that of qPCR (4.4%). CONCLUSIONS: This study successfully developed a sensitive, specific and repeatable ddPCR assay for PCV4 detection, which can be widely used in clinical diagnosis of PCV4 infections.

摘要

背景：猪圆环病毒 4 型（PCV4）是一种新出现的病毒，于 2019 年首次发现，可能对养猪业构成潜在威胁。液滴数字 PCR（ddPCR）是一种绝对定量方法，具有高灵敏度和准确性。本研究开发了一种检测 PCV4 的新型 ddPCR 检测方法。此外，我们评估了 ddPCR 和 TaqMan 实时定量 PCR（qPCR）的检测限、灵敏度、特异性和重现性，并测试了 160 个临床样本，以比较两种方法的检测率。

结果：ddPCR 的检测限为 0.54 拷贝/μL，比 qPCR 高 10.6 倍。ddPCR 和 qPCR 检测均表现出良好的线性和重复性，建立的 ddPCR 方法对 PCV4 具有高度特异性。临床样本检测结果表明，ddPCR 的阳性率（5.6%）高于 qPCR（4.4%）。

结论：本研究成功开发了一种用于检测 PCV4 的灵敏、特异和可重复的 ddPCR 检测方法，可广泛应用于 PCV4 感染的临床诊断。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/1fcfbce81c34/12917_2023_3690_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/a1aa7880ea0f/12917_2023_3690_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/82b0dad2e6d3/12917_2023_3690_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/633102973dcc/12917_2023_3690_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/1fcfbce81c34/12917_2023_3690_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/a1aa7880ea0f/12917_2023_3690_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/82b0dad2e6d3/12917_2023_3690_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/633102973dcc/12917_2023_3690_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9bc9/10464377/1fcfbce81c34/12917_2023_3690_Fig4_HTML.jpg

相似文献

[1]
Development of a droplet digital PCR method for detection of porcine circovirus 4.

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[2]
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[3]
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[4]
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[5]
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[6]
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[7]
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[8]
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[9]
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[10]
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引用本文的文献

[1]
Development of a duplex real-time recombinase aided amplification assay for the simultaneous and rapid detection of PCV3 and PCV4.

Virol J. 2025-2-1

[2]
Detection of Porcine Circovirus (PCV) Using CRISPR-Cas12a/13a Coupled with Isothermal Amplification.

Viruses. 2024-9-30

[3]
Droplet digital PCR aids in the diagnosis of children with fever of unknown origin --A typical case report.

Heliyon. 2024-5-9

本文引用的文献

[1]
Rapid detection of porcine circovirus type 4 multienzyme isothermal rapid amplification.

Front Vet Sci. 2022-7-28

[2]
Development of a Real-time TaqMan PCR Assay for The Detection of Porcine Circovirus 4.

J Vet Res. 2022-3-25

[3]
Porcine circovirus 4 rescued from an infectious clone is replicable and pathogenic in vivo.

Transbound Emerg Dis. 2022-9

[4]
Development of a one-step multiplex qRT-PCR assay for the detection of African swine fever virus, classical swine fever virus and atypical porcine pestivirus.

BMC Vet Res. 2022-1-18

[5]
Detection of a novel porcine circovirus 4 in Korean pig herds using a loop-mediated isothermal amplification assay.

J Virol Methods. 2022-1

[6]
Development and application of an indirect ELISA for the detection of antibody to porcine circovirus 4 in pigs.

Transbound Emerg Dis. 2021-11

[7]
Phylogenetic analysis of porcine circovirus 4 in Henan Province of China: A retrospective study from 2011 to 2021.

Transbound Emerg Dis. 2022-7

[8]
Retrospective surveillance of porcine circovirus 4 in pigs in Inner Mongolia, China, from 2016 to 2018.

Arch Virol. 2021-7

[9]
Detection and genetic characterization of porcine circovirus 4 (PCV4) in Guangxi, China.

Gene. 2021-3-20

[10]
Development and application of a quadruplex real-time PCR assay for differential detection of porcine circoviruses (PCV1 to PCV4) in Jiangsu province of China from 2016 to 2020.

Transbound Emerg Dis. 2021-5

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