Development of a real-time PCR assay for the detection and identification of Staphylococcus capitis, Staphylococcus haemolyticus and Staphylococcus warneri
Development of a real-time PCR assay for the detection and identification of Staphylococcus capitis, Staphylococcus haemolyticus and Staphylococcus warneri
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DOI：
10.1099/jmm.0.47235-0
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发表时间：
2007-10-01
影响因子：
3
通讯作者：
Masuda, Shogo
中科院分区：
文献类型：
--
作者：
Iwase, Tadayuki;Seki, Keiko;Masuda, Shogo
头状葡萄球菌、溶血葡萄球菌和瓦梅里葡萄球菌是凝固酶阴性葡萄球菌。每个物种都有不同的特征，受损宿主的病理学也存在差异。因此，开发一种特异性简单的检测方法来鉴定这些葡萄球菌非常重要。在此，报道了一种针对这些细菌的超氧化物歧化酶 A 编码基因的物种特异性实时 PCR 检测。引物设计有与其他细菌不互补的碱基。该碱基位于引物（3' 错配引物）的 3' 末端，具有高特异性。然后使用实时 PCR 评估这些引物。它们只与目标细菌发生反应。此外，当使用从不同数量的葡萄球菌细胞（10(1)-10(7)细胞）提取的基因组DNA进行实验时，观察到稳定的定量反应。这些结果表明该方法可用于头状链球菌、溶血链球菌和华氏链球菌的鉴定和定量分析。
Staphylococcus capitis, Staphylococcus haemolyticus and Staphylococcus wameri are coagulase-negative staphylococci. Each species has different characteristics, and a difference in pathology is also seen in compromised hosts. Therefore, the development of a species-specific simple detection method for the identification of these staphylococci is important. Here, a species-specific real-time PCR assay is reported that targets the superoxide dismutase A-encoding gene of these bacteria. Primers were designed with a base that was non-complementary with regard to the other bacteria. This base was at the 3' end of the primer (3' mismatch primer) and conferred high specificity. These primers were then evaluated using real-time PCR. They reacted only with the target bacterium. In addition, stable quantitative reactions were observed when experiments were performed using genomic DNA extracted from varying numbers of staphylococci cells (10(1)-10(7) cells). These results indicate that this method is useful for the identification and quantitative analysis of S. capitis, S. haemolyticus and S. warneri.