High specific MNase assay for rapid identification of Staphylococcus aureus using AT-rich dsDNA substrate
High specific MNase assay for rapid identification of Staphylococcus aureus using AT-rich dsDNA substrate
复制标题
使用富含 AT 的 dsDNA 底物快速鉴定金黄色葡萄球菌的高特异性 MNase 测定
DOI：
10.1016/j.talanta.2019.06.029
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发表时间：
2019
期刊：
影响因子：
6.1
通讯作者：
Feng Bo
中科院分区：
文献类型：
--
作者：
Wang Xuan;Long Caicheng;Xu Xiaoyan;Qing Taiping;Zhang Peng;Feng Bo
Micrococcal nuclease (MNase) is a nonspecific endo-exonuclease that digests single-stranded/double-stranded DNA and RNA. The existence of MNase can serves as an important diagnostic biomarker of Staphylococcus aureus (S. aureus) infection. However, most of the substrates in MNase-based sensors are single-stranded DNA, which could also be digested by exonuclease I or S1 nuclease and interfere the MNase detection. In this work, we developed a highly selective fluorescent method for MNase detection using a specific dsDNA and nucleic acid dye SYBR Green I (SGI) as the indicator. After rational design, an AT-rich dsDNA with 3′ protruding termini was screened as the high-specific substrate of MNase assay and efficient enhancer of SGI. The AT-rich dsDNA substrate can resist the digestion of other exonuclease and greatly enhance the fluorescence of SGI. This high-specific substrate-based probe can detect MNase in buffer as well as biological sample with highly selectivity. Moreover, this method was also applied to monitor the MNase secreted byS. aureus. Thus, the proposed MNase-based assay has a strong potential to identifyS. aureusin food safety and microbial infection due to its excellent analytical sensitivity and high selectivity.
