Integrated Detection of Extended-Spectrum-Beta-Lactam Resistance by DNA Microarray-Based Genotyping of TEM, SHV, and CTX-M Genes
Integrated Detection of Extended-Spectrum-Beta-Lactam Resistance by DNA Microarray-Based Genotyping of TEM, SHV, and CTX-M Genes
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DOI：
10.1128/jcm.00765-09
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发表时间：
2010-02-01
影响因子：
9.4
通讯作者：
Bachmann, Till T.
中科院分区：
文献类型：
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作者：
Leinberger, Dirk M.;Grimm, Verena;Bachmann, Till T.
TEM、SHV 或 CTX-M 型超广谱 β-内酰胺酶 (ESBL) 赋予革兰氏阴性菌对 β-内酰胺抗生素的耐药性。这些酶对β-内酰胺抗生素的活性及其对抑制剂的耐药性可能受到单核苷酸水平遗传变异的影响。在这里，我们描述了寡核苷酸微阵列的开发和验证，用于通过同时对 bla(TEM)、bla(SHV) 和 bla(CTX-M) 进行基因分型来快速鉴定革兰氏阴性菌中的 ESBL。该阵列由 618 个探针组成，涵盖负责 156 个氨基酸取代的突变。由于这包括前所未有的基因分型覆盖范围，ESBL 芯片在流行病学研究和感染控制方面具有很高的潜力。 ESBL 微阵列的检测时间为 5 小时，也可能成为未来快速抗菌药物耐药性测试开发的一个有吸引力的选择。 DNA 微阵列的有效性通过在临床常规过程中收集的 60 个盲法临床分离株得到证明。其中 58 人的表型特征为 ESBL 生产者。该芯片的特征在于其分辨率、表型-基因型相关性以及解析混合基因型的能力。 ESBL 表型可以正确归因于 bla(CTX-M) (76%)、bla(SHV) (22%) 或两者 (2%) 的 ESBL 变体，而没有发现 bla(TEM) 的 ESBL 变体。最常见的 ESBL 是 CTX-M-15 (57%) 和 SHV-12 (18%)。
Extended-spectrum beta-lactamases (ESBL) of the TEM, SHV, or CTX-M type confer resistance to beta-lactam antibiotics in Gram-negative bacteria. The activity of these enzymes against beta-lactam antibiotics and their resistance against inhibitors can be influenced by genetic variation at the single-nucleotide level. Here, we describe the development and validation of an oligonucleotide microarray for the rapid identification of ESBLs in Gram-negative bacteria by simultaneously genotyping bla(TEM), bla(SHV), and bla(CTX-M). The array consists of 618 probes that cover mutations responsible for 156 amino acid substitutions. As this comprises unprecedented genotyping coverage, the ESBL array has a high potential for epidemiological studies and infection control. With an assay time of 5 h, the ESBL microarray also could be an attractive option for the development of rapid antimicrobial resistance tests in the future. The validity of the DNA microarray was demonstrated with 60 blinded clinical isolates, which were collected during clinical routines. Fifty-eight of them were characterized phenotypically as ESBL producers. The chip was characterized with regard to its resolution, phenotype-genotype correlation, and ability to resolve mixed genotypes. ESBL phenotypes could be correctly ascribed to ESBL variants of bla(CTX-M) (76%), bla(SHV) (22%), or both (2%), whereas no ESBL variant of bla(TEM) was found. The most prevalent ESBLs identified were CTX-M-15 (57%) and SHV-12 (18%).