制备和应用多克隆抗体快速检测放线菌绿环斑相关病毒。

IF 3.8 3区 医学 Q2 VIROLOGY
Viruses-Basel Pub Date : 2024-10-11 DOI:10.3390/v16101600
Jing Shang, Hongping Feng, Yuxuan Wang, Yunan Wang, Xiao Zhang, Zhouyu Zhang
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引用次数: 0

摘要

猕猴桃叶枯环斑病相关病毒(AcCRaV,Emaravirus actinidiae)在中国猕猴桃中流行,导致大量减产。该病毒的症状错综复杂,给诊断带来了挑战,因此需要一种快速、准确的检测方法来进行有效控制。本研究从四川省猕猴桃主要产区收集并分析了 AcCRaV 分离物,并选择具有代表性的菌株作为实验材料。设计了针对 AcCRaV 核蛋白基因的引物,并对其密码子用法进行了优化,以提高其性能。利用多克隆抗体开发了多种血清学方法,包括酶联免疫吸附试验、点免疫结合试验和 AcCRaV 特异性金免疫层析带(AcCRaV-GICS)。使用这些方法检测野外样本时,均显示出较高的特异性和灵敏度。此外,从大量野外样本中获得的结果与 RT-PCR 分析得出的结果一致,进一步验证了我们方法的适用性。目前还缺乏一种能够处理大量感染 AcCRaV 的野外样本的检测方法;因此,我们的系统构建为弥补这一空白提供了重要参考。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Preparation and Application of Polyclonal Antibodies for the Rapid Detection of Actinidia Chlorotic Ringspot-Associated Virus.

Actinidia chlorotic ringspot-associated virus (AcCRaV, Emaravirus actinidiae) is prevalent in Chinese kiwifruit, leading to substantial yield reduction. The intricate nature of symptoms presents diagnostic challenges, underscoring the necessity for a rapid and accurate detection method that facilitates effective control. In this investigation, AcCRaV isolates from key kiwi-producing regions in Sichuan province were collected and analyzed, with representative strains chosen as experimental materials. Primers targeting the nucleoprotein gene of AcCRaV were designed, and their codon usage was optimized to enhance performance. Various serological methods utilizing polyclonal antibodies were developed, including ELISA, dot immunobinding assay, and AcCRaV-specific gold immunochromatographic bands (AcCRaV-GICS). Field samples exhibited high specificity and sensitivity when tested using these methods. Furthermore, the results obtained from a large number of field samples are consistent with those derived from RT-PCR analysis, further validating the applicability of our approach. A detection method capable of handling a large volume of field samples infected with AcCRaV is currently lacking; thus, our system construction provides an important reference for addressing this gap.

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来源期刊
Viruses-Basel
Viruses-Basel VIROLOGY-
CiteScore
7.30
自引率
12.80%
发文量
2445
审稿时长
1 months
期刊介绍: Viruses (ISSN 1999-4915) is an open access journal which provides an advanced forum for studies of viruses. It publishes reviews, regular research papers, communications, conference reports and short notes. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced. We also encourage the publication of timely reviews and commentaries on topics of interest to the virology community and feature highlights from the virology literature in the ''News and Views'' section. Electronic files or software regarding the full details of the calculation and experimental procedure, if unable to be published in a normal way, can be deposited as supplementary material.
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