建立基于内切酶活性的犬细小病毒快速恒温检测方法。

IF 3.7 2区 生物学 Q2 MICROBIOLOGY
Microbiology spectrum Pub Date : 2024-11-05 Epub Date: 2024-09-17 DOI:10.1128/spectrum.04222-23
Shaoting Weng, Shengming Ma, Yueteng Xing, Wenhui Zhang, Yinrong Wu, Mengyao Fu, Zhongyi Luo, Qiuying Li, Sen Lin, Longfei Zhang, Yao Wang
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引用次数: 0

摘要

犬细小病毒 (CPV) 可导致幼犬的高发病率和高死亡率,对宠物犬和饲养业都构成重大威胁。快速、准确、方便的检测方法对于犬细小病毒的早期干预和治疗非常重要。在这项研究中,我们提出了一种名为核酸错配酶消化(NMED)的可视化 CPV 检测系统。该系统结合了环介导等温扩增(LAMP)、用于基因错配检测的内切酶和胶体金横向层析技术。我们证明,NMED 可以诱导样本中的扩增子与特异性标记探针结合,进而引发内切酶的消化。通过将内切酶和胶体金横向层析相结合,并辅以简单的温控装置,提高了 LAMP 的灵敏度和可视性。NMED 检测的灵敏度为 1 拷贝/μL,与定量 PCR(qPCR)一致。该方法在 20 份可能感染 CPV 的临床样本中进行了验证,评估了 15 份阳性样本和 5 份阴性样本,其检测准确度与 qPCR 一致。作为一种快速、准确、便捷的分子诊断方法,NMED 在病原微生物检测领域具有巨大的应用潜力:重要意义:NMED 方法已在实验室建立并用于 CPV 检测。该方法具有取样简单、灵敏度高、结果直观、无需昂贵设备等优点。该方法的建立具有商业潜力,并为未来病原微生物临床检测的发展提供了一种新的方法和理念。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Toward establishing a rapid constant temperature detection method for canine parvovirus based on endonuclease activities.

Canine parvovirus (CPV) can cause high morbidity and mortality rates in puppies, posing a significant threat to both pet dogs and the breeding industry. Rapid, accurate, and convenient detection methods are important for the early intervention and treatment of canine parvovirus. In this study, we propose a visual CPV detection system called nucleic acid mismatch enzyme digestion (NMED). This system combines loop-mediated isothermal amplification (LAMP), endonuclease for gene mismatch detection, and colloidal gold lateral chromatography. We demonstrated that NMED can induce the binding of the amplicon from the sample to the specific labeling probe, which in turn triggers digestion by the endonuclease. The sensitivity and visual visibility of LAMP were increased by combining endonuclease and colloidal gold lateral chromatography assisted by a simple temperature-controlled device. The sensitivity of the NMED assay was 1 copy/μL, which was consistent with quantitative PCR (qPCR). The method was validated with 20 clinical samples that potentially had CPV infection; 15 positive samples and 5 negative samples were evaluated; and the detection accuracy was consistent with that of qPCR. As a rapid, accurate, and convenient molecular diagnostic method, NMED has great potential for application in the field of pathogenic microorganism detection.

Importance: The NMED method has been established in the laboratory and used for CPV detection. The method has several advantages, including simple sampling, high sensitivity, intuitive results, and no requirement for expensive equipment. The establishment of this method has commercial potential and offers a novel approach and concept for the future development of clinical detection of pathogenic microorganisms.

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来源期刊
Microbiology spectrum
Microbiology spectrum Biochemistry, Genetics and Molecular Biology-Genetics
CiteScore
3.20
自引率
5.40%
发文量
1800
期刊介绍: Microbiology Spectrum publishes commissioned review articles on topics in microbiology representing ten content areas: Archaea; Food Microbiology; Bacterial Genetics, Cell Biology, and Physiology; Clinical Microbiology; Environmental Microbiology and Ecology; Eukaryotic Microbes; Genomics, Computational, and Synthetic Microbiology; Immunology; Pathogenesis; and Virology. Reviews are interrelated, with each review linking to other related content. A large board of Microbiology Spectrum editors aids in the development of topics for potential reviews and in the identification of an editor, or editors, who shepherd each collection.
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