一种使用 Surveyor 核酸内切酶检测烟曲霉基因突变的简单方法。
A Simple Method To Detect Point Mutations in Aspergillus fumigatus Gene Using a Surveyor Nuclease Assay.
机构信息
Medical Mycology Research Center, Chiba University, Chiba, Japan.
Department of Respiratory Medicine, Tokyo Medical and Dental University, Tokyo, Japan.
出版信息
Antimicrob Agents Chemother. 2020 Mar 24;64(4). doi: 10.1128/AAC.02271-19.
Abstract
One of the main mechanisms of azole resistance of is thought to be a reduction in the drug's affinity for the target molecule, Cyp51A, due to its amino acid mutation(s). It is known that the azole resistance pattern is closely related to the mutation site(s) of the molecule. In this study, we tried to develop a simple and rapid detection method for mutations using the endonuclease Surveyor nuclease. The Surveyor nuclease assay was verified using several azole-resistant strains of that possess point mutations in Cyp51A. For validation of the Surveyor nuclease assay, blind tests were conducted using 48 strains of (17 azole-resistant and 31 azole-susceptible strains). The Surveyor nuclease assay could rapidly detect mutations with one primer set. Also, all the tested strains harboring different single point mutations could be clearly distinguished from the wild type. The Surveyor nuclease assay is a simple method that can detect mutations rapidly.
摘要
一种被认为是唑类耐药的主要机制是由于其氨基酸突变导致药物对靶分子 Cyp51A 的亲和力降低。已知唑类耐药模式与分子的突变部位密切相关。在这项研究中，我们试图使用内切酶 Surveyor 核酸酶开发一种简单快速的检测方法。使用几种在 Cyp51A 中具有点突变的唑类耐药的 进行了 Surveyor 核酸酶测定的验证。为了验证 Surveyor 核酸酶测定，使用 48 株 （17 株唑类耐药株和 31 株唑类敏感株）进行了盲法测试。Surveyor 核酸酶测定可以使用一组引物快速检测 突变。此外，所有测试的株系携带不同的 单点突变都可以与野生型明显区分开来。Surveyor 核酸酶测定是一种快速检测 突变的简单方法。
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本文引用的文献
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J Mol Diagn. 2019 Mar;21(2):286-295. doi: 10.1016/j.jmoldx.2018.10.004. Epub 2018 Dec 4.
2
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3
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4
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9
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Theriogenology. 2016 Nov;86(8):1886-1896.e1. doi: 10.1016/j.theriogenology.2016.06.010. Epub 2016 Jun 15.
10
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