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稻曲病间接 ELISA 和 Dot-Blot 检测方法的建立。

Development of an Indirect ELISA and Dot-Blot Assay for Serological Detection of Rice Tungro Disease.

机构信息

Institute of Health & Community Medicine, Universiti Malaysia Sarawak, 94300 Kota Samarahan, Sarawak, Malaysia.

Agriculture Research Centre (ARC), Department of Agriculture (DOA) Sarawak, Semongok, Borneo Heights Road, 93250 Kuching, Sarawak, Malaysia.

出版信息

Biomed Res Int. 2017;2017:3608042. doi: 10.1155/2017/3608042. Epub 2017 Oct 22.

Abstract

Rice tungro disease (RTD) is one of the most destructive diseases of rice in South and Southeast Asia. RTD is routinely detected based on visual observation of the plant. However, it is not always easy to identify the disease in the field as it is often confused with other diseases or physiological disorders. Here we report the development of two serological based assays for ease of detection of RTD. In this study we had developed and optimized an indirect ELISA and dot-blot assay for detection of RTD. The efficiency of both assays was evaluated by comparing the specificity and sensitivity of the assays to PCR assay using established primer sets. The indirect ELISA showed 97.5% and 96.6%, while the dot-blot assay showed 97.5% and 86.4% sensitivity and specificity, respectively, when compared to established PCR method. The high sensitivity and specificity of the two assays merit the use of both assays as alternative methods to diagnose RTD. Furthermore, the dot-blot assay is a simple, robust, and rapid diagnostic assay that is suitable for field test for it does not require any specialized equipment. This is a great advantage for diagnosing RTD in paddy fields, especially in the rural areas.

摘要

水稻矮缩病(RTD)是南亚和东南亚地区水稻最具破坏性的疾病之一。RTD 的常规检测是基于对植物的视觉观察。然而,由于它经常与其他疾病或生理障碍混淆,因此在田间并不总是容易识别。在这里,我们报告了两种基于血清学的检测方法的开发,以方便检测 RTD。在本研究中,我们开发并优化了间接 ELISA 和斑点印迹分析检测 RTD。通过将两种检测方法与使用既定引物组的 PCR 检测方法进行比较,评估了这两种检测方法的特异性和敏感性。间接 ELISA 的敏感性和特异性分别为 97.5%和 96.6%,而斑点印迹分析的敏感性和特异性分别为 97.5%和 86.4%。两种检测方法的高敏感性和特异性使得这两种检测方法都可以作为替代方法来诊断 RTD。此外,斑点印迹分析是一种简单、稳健、快速的诊断检测方法,不需要任何特殊设备,非常适合现场测试。这对于在稻田中诊断 RTD 特别是在农村地区是一个很大的优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/db89/5671674/ca7a36ad0840/BMRI2017-3608042.001.jpg

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