Center for Nano and Material Sciences, Jain University, Jain Global campus, Bengaluru, Karnataka 562112, India.
Analyst. 2019 Jun 24;144(13):3999-4005. doi: 10.1039/c8an02116c.
Micrococcal nuclease (MNase) is an extracellular endonuclease of Staphylococcus aureus (S. aureus). It digests single stranded nucleic acid. The presence of MNase is the gold standard to identify S. aureus and its content. The present study reports the ultrahigh sensitive and selective fluorescence platform for MNase detection, designed and developed based on the surface energy transfer mechanism. A "proof of concept" is being developed based on monoclonal antibody-conjugated quantum dots (mAb-QDs), wherein mAb-QDs act as donors and graphene oxide (GO) acts as an acceptor. mAb-QDs in close proximity to GO undergo adsorption due to weak affinity between them and this results in fluorescence quenching by the transfer of surface energy from mAb-QDs to GO. During sensing, a much stronger affinity of mAb-QDs towards MNase inhibits the energy transfer to GO and this allows the regaining of fluorescence. Immobilized mAb-QDs on nitrocellulose membrane strips were fabricated and tested for "ON-OFF-ON" sensing of MNase. The limit of detection for fluorescence based assay and strips is found to be 0.3 ng mL-1 and 0.5 ng mL-1, respectively. The developed strips were applied on real samples for the detection of S. aureus.
微球菌核酸酶(MNase)是金黄色葡萄球菌(S. aureus)的一种细胞外内切酶。它可消化单链核酸。MNase 的存在是鉴定金黄色葡萄球菌及其含量的金标准。本研究报告了一种基于表面能量转移机制设计和开发的超高灵敏和选择性 MNase 检测荧光平台。正在基于单克隆抗体偶联量子点(mAb-QDs)开发“概念验证”,其中 mAb-QDs 作为供体,氧化石墨烯(GO)作为受体。由于它们之间的亲和力较弱,因此靠近 GO 的 mAb-QDs 会因吸附而发生,这导致表面能量从 mAb-QDs 转移到 GO 导致荧光猝灭。在传感过程中,mAb-QDs 对 MNase 的亲和力要强得多,从而抑制了向 GO 的能量转移,从而恢复了荧光。在硝酸纤维素膜条上固定化 mAb-QDs 用于 MNase 的“开-关-开”感应。荧光测定法和条带的检测限分别为 0.3 ng mL-1 和 0.5 ng mL-1。开发的条带已应用于实际样品中以检测金黄色葡萄球菌。