Development of TaqMan Probe-Based Real-Time PCR Method for erm(A), erm(B), and erm(C), Rapid Detection of Macrolide-Lincosamide-Streptogramin B Resistance Genes, from Clinical Isolates
Development of TaqMan Probe-Based Real-Time PCR Method for erm(A), erm(B), and erm(C), Rapid Detection of Macrolide-Lincosamide-Streptogramin B Resistance Genes, from Clinical Isolates
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DOI：
10.4014/jmb.0902.0062
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发表时间：
2009-11-01
影响因子：
2.8
通讯作者：
Choi, Sung-Sook
中科院分区：
文献类型：
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作者：
Jung, Jae-Hyuk;Yoon, Eun-Jeong;Choi, Sung-Sook
为实现对大环内酯类-林可酰胺类-链阳性菌素B耐药基因的准确、快速检测，我们建立了一种基于TaqMan探针的实时荧光定量PCR（Q-PCR）方法，并与目前应用最广泛的PCR方法进行了比较。Q-PCR的检出限为5 fg基因组DNA或5-8 CFU的金黄色葡萄球菌细菌细胞。Q-PCR的应用可将检测时间从3-4 h缩短到50 min左右，表明Q-PCR不仅具有较高的敏感性和特异性，而且是最快速的诊断方法。因此，Q-PCR检测的适当应用将能够快速、准确地识别临床和其他样本中的erm基因。
To achieve more accurate and rapid detection of macrolide-lincosamide-streptogramin B resistance genes, erm(A), erm(B), and erm(C), we developed a TaqMan probe-based real-time PCR (Q-PCR) method and compared it with conventional PCR (C-PCR), which is the most widely using erm gene identification method. The detection limit of Q-PCR was 5 fg of genomic DNA or 5-8 CFU of bacterial cells of Staphylococcus aureus. The utilization of Q-PCR might shorten the time to erm detection from 3-4 h to about 50 min. These data indicated that Q-PCR assay appears to be not only highly sensitive and specific, but also the most rapid diagnostic method. Therefore, the appropriate application of the Q-PCR assay will permit rapid and accurate identification of erm genes from clinical and other samples.