Real-time PCR methods for quantitative monitoring of streptomycin and tetracycline resistance genes in agricultural ecosystems

被引：72

作者：

Walsh, F. ^{[1
]}

Ingenfeld, A. ^{[1
]}

Zampicolli, M. ^{[1
]}

Hilber-Bodmer, M. ^{[1
]}

Frey, J. E. ^{[1
]}

Duffy, B. ^{[1
]}

机构：

[1] ACW, Div Plant Protect, CH-8820 Wadenswil, Switzerland

来源：

JOURNAL OF MICROBIOLOGICAL METHODS | 2011年 / 86卷 / 02期

关键词：

Antibiotic resistance; Streptomycin; Tetracycline; Agriculture; qPCR; TaqMan (R); BACTERIAL ANTIBIOTIC-RESISTANCE; STAPHYLOCOCCUS-AUREUS; MANURE; QUANTIFICATION; ABUNDANCE; LAGOONS; SOIL; EXPRESSION; DIVERSITY; HABITATS;

D O I：

10.1016/j.mimet.2011.04.011

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：150 / 155

页数：6

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