Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus
Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus
复制标题
DOI：
10.1128/jcm.41.9.4089-4094.2003
复制
发表时间：
2003-09-01
影响因子：
9.4
通讯作者：
Witte, W
中科院分区：
文献类型：
--
作者：
Strommenger, B;Kettlitz, C;Witte, W
在这项研究中，我们描述了一种多重PCR方法来检测九种临床相关的金黄色葡萄球菌耐药基因。优化了同时扩增编码甲氧西林耐药基因mecA、氨基糖苷耐药基因AACA-APHD、四环素耐药基因etK、四环素耐药基因etM、耐大环内酯类药物林可西林-链霉素B的基因erm(C)、耐大环内酯类药物链霉素B的基因vat(A)、耐药基因vat(B)和耐链球菌素A的基因vat(C)的扩增条件。另加一对扩增葡萄球菌16S rDNA片段的引物作为阳性对照。对30株不同的金黄色葡萄球菌进行了多重PCR检测，其结果与肉汤微量稀释法获得的表型耐药性数据相一致。多重聚合酶链式反应提供了一种快速、简便和准确的抗生素耐药谱鉴定方法，可用于临床诊断以及在流行病学研究中监测抗生素耐药性决定因素的传播。
In this study we describe a multiplex PCR assay for the detection of nine clinically relevant antibiotic resistance genes of Staphylococcus aureus. Conditions were optimized to amplify fragments of mecA (encoding methicillin resistance), aacA-aphD (aminoglycoside resistance), tetK, tetM (tetracycline resistance), erm(A), erm(C) (macrolide-lincosamide-streptogramin B resistance), vat(A), vat(B), and vat(C) (streptogramin A resistance) simultaneously in one PCR amplification. An additional primer pair for the amplification of a fragment of the staphylococcal 16S rDNA was included as a positive control. The multiplex PCR assay was evaluated on 30 different S. aureus isolates, and the PCR results correlated with the phenotypic antibiotic resistance data obtained by the broth microdilution assay. The multiplex PCR assay offers a rapid, simple, and accurate identification of antibiotic resistance profiles and could be used in clinical diagnosis as well as for the surveillance of the spread of antibiotic resistance determinants in epidemiological studies.