Real-time PCR methods for quantitative monitoring of streptomycin and tetracycline resistance genes in agricultural ecosystems
Real-time PCR methods for quantitative monitoring of streptomycin and tetracycline resistance genes in agricultural ecosystems
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DOI：
10.1016/j.mimet.2011.04.011
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发表时间：
2011-08-01
影响因子：
2.2
通讯作者：
Duffy, B.
中科院分区：
文献类型：
--
作者：
Walsh, F.;Ingenfeld, A.;Duffy, B.
抗生素在植物农业中的应用主要用于控制仁果果园中由梨火疫病菌引起的火疫病。为了促进抗生素应用的环境影响评估，我们开发并验证了植物和土壤样品中链霉素（strA，strB，aadA和插入序列IS1133）和四环素（tetB，tetM和tetW）抗性元素的非培养型定量实时PCR多重检测。无论是直接从接种细菌的细菌、植物和土壤样本还是粪便浆处理之前和之后的土壤样本中提取DNA，qPCR都是可重复且一致的。在泥浆处理前后的土壤中最常鉴定的基因是strB、aadA、tetB和tetM。所有测试的基因在土壤中检测到预泥浆处理，和减少tetB和链霉素抗性基因的相对浓度观察泥浆处理后采取的样品。这些多重qPCR检测为复杂环境样品基质中抗生素耐药基因的同时定量提供了一种具有成本效益的可靠方法。(C)2011爱思唯尔有限公司版权所有。
Antibiotic application in plant agriculture is primarily used to control fire blight caused by Erwinia amylovora in pome fruit orchards. In order to facilitate environmental impact assessment for antibiotic applications, we developed and validated culture-independent quantitative real-time PCR multiplex assays for streptomycin (strA, strB, aadA and insertion sequence IS1133) and tetracycline (tetB, tetM and tetW) resistance elements in plant and soil samples. The qPCR were reproducible and consistent whether the DNA was extracted directly from bacteria, plant and soil samples inoculated with bacteria or soil samples prior to and after manure slurry treatment. The genes most frequently identified in soils pre- and post-slurry treatment were strB, aadA, tetB and tetM. All genes tested were detected in soils pre-slurry treatment, and a decrease in relative concentrations of tetB and the streptomycin resistance genes was observed in samples taken post-slurry treatment. These multiplex qPCR assays offer a cost-effective, reliable method for simultaneous quantification of antibiotic resistance genes in complex, environmental sample matrices. (C) 2011 Elsevier B.V. All rights reserved.