Detection of Streptococcus pyogenes using rapid visual molecular assay
Detection of Streptococcus pyogenes using rapid visual molecular assay
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使用快速可视化分子测定法检测化脓性链球菌
DOI：
10.1093/femsle/fnv148
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发表时间：
2015-09-01
影响因子：
2.1
通讯作者：
Yuan, Jing
中科院分区：
文献类型：
--
作者：
Zhao, Xiangna;He, Xiaoming;Yuan, Jing
化脓性链球菌在世界许多地区是一种日益重要的病原体。快速准确地检测化脓性葡萄球菌有助于控制感染。本研究建立了一种环介导的等温扩增（LAMP）方法，并验证了该方法对化脓性葡萄球菌的特异性检测。该分析包含两种方法：使用钙黄蛋白/Mn2+复合物的显色分析和实时浊度监测来评估反应。两种方法均在64℃等温条件下60 min内检测到目标DNA。实验采用专门设计的引物以spy1258为靶点，正确鉴定出111株化脓性链球菌和32株非化脓性链球菌。化脓性菌株，包括其他种类的链球菌属。采用参考菌株进行的试验表明，LAMP法具有很高的特异性。检测限为1.49 pg DNA，灵敏度是PCR的10倍。本研究建立的LAMP检测方法简单、快速、灵敏，不依赖于任何特殊设备；因此，它可以用于临床诊断。
Streptococcus pyogenes is an increasingly important pathogen in many parts of the world. Rapid and accurate detection of S. pyogenes aids in the control of the infection. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the specific detection of S. pyogenes. The assay incorporates two methods: a chromogenic analysis using a calcein/Mn2+ complex and real-time turbidity monitoring to assess the reaction. Both methods detected the target DNA within 60 min under 64 degrees C isothermal conditions. The assay used specifically designed primers to target spy1258, and correctly identified 111 strains of S. pyogenes and 32 non-S. pyogenes strains, including other species of the genus Streptococcus. Tests using reference strains showed that the LAMP assay was highly specific. The sensitivity of the assay, with a detection limit of 1.49 pg DNA, was 10-fold greater than that of PCR. The LAMP assay established in this study is simple, fast and sensitive, and does not rely upon any special equipment; thus, it could be employed in clinical diagnosis.