Detection of antibiotic resistance genes of Escherichia coli from domestic livestock in south east Nigeria with DNA microarray

Chijioke A. Nsofor, Christian U. Iroegbu Douglas R. Call, M. Davies
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引用次数: 5

Abstract

DNA microarray was developed for detection of up to 90 antibiotic resistance genes in Escherichia coli by hybridization. Each antibiotic resistance gene was represented by two specific oligonucleotides chosen from consensus sequences of gene families. A total of 203 oligonucleotides (50-100 base) were spotted onto the microarray. The sequence identity of each gene was compared with GenBank sequen-ces, biotin was used as the positive control and 16s rRNA as orientation. Of the 40 E. coli isolates analyzed in this study, 37 were identified as having, at least, one antibiotic resistance gene. Among the different antibiotic resistance genes detected, bla-CMY-2 and strA were the most prevalent occurring in 28 (70%) of the isolates, respectively. Other common genes included were TEM1 11(27.5%), Sul2 14 (35%) and TetA 21(52.5%). The microarray genotyping corresponded with the phenotype of the strains. The disposable microarray presents the advantage of rapidly screening bacteria for the pre-sence of known antibiotic resistance genes. This technology has a large potential for applications in basic research, food safety, and surveillance programs for antimicrobial resistance.   Key words: DNA microarray, antibiotic resistance, Escherichia coli.
DNA芯片技术检测尼日利亚东南部家畜大肠杆菌抗生素耐药基因
利用DNA微阵列技术对大肠杆菌中多达90个耐药基因进行杂交检测。每个抗生素耐药基因由从基因家族一致序列中选择的两个特定寡核苷酸表示。共有203个寡核苷酸(50-100个碱基)被标记到芯片上。以生物素为阳性对照,以16s rRNA为定向,与GenBank序列进行序列一致性比较。在本研究分析的40株大肠杆菌中,有37株被鉴定为至少具有一种抗生素耐药基因。在检测到的不同耐药基因中,bla-CMY-2和strA最常见,分别出现在28株(70%)菌株中。其他常见基因包括tem111(27.5%)、sul214(35%)和teta21(52.5%)。微阵列基因分型与菌株表型一致。一次性微阵列具有快速筛选已知抗生素耐药基因存在的细菌的优势。这项技术在基础研究、食品安全和抗菌素耐药性监测规划方面具有巨大的应用潜力。关键词:DNA芯片,抗生素耐药性,大肠杆菌
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