A Bioluminescent Sensor for Rapid Detection of PPEP-1, a Clostridioides difficile Biomarker.
A Bioluminescent Sensor for Rapid Detection of PPEP-1, a Clostridioides difficile Biomarker.
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DOI：
10.3390/s21227485
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发表时间：
2021-11-11
期刊：
影响因子：
--
通讯作者：
Prescher JA
中科院分区：
文献类型：
--
作者：
Ng KK;Reinert ZE;Corver J;Resurreccion D;Hensbergen PJ;Prescher JA
目前在非医院环境中对艰难梭菌的检测是外包的且耗时，导致感染个体的诊断和隔离被延迟。我们设计了一种更快速的现场检测，其特点是“开启”艰难梭菌特异性蛋白酶 PPEP-1 的生物发光读数。 NanoLuc 是一种明亮且稳定的荧光素酶，被 PPEP-1 响应肽尾“囚禁”，抑制发光。蛋白水解后，肽被释放，NanoLuc 活性恢复，提供可见的读数。生物发光传感器检测到 PPEP-1 浓度低至 10 nM。几分钟内就实现了传感器的解锁，并使用数码相机捕获了信号。重要的是，该传感器在环境温度下也能发挥作用，并且与粪便材料兼容，这表明它可以轻松部署在各种环境中。
Current assays for Clostridioides difficile in nonhospital settings are outsourced and time-intensive, resulting in both delayed diagnosis and quarantining of infected individuals. We designed a more rapid point-of-care assay featuring a “turn-on” bioluminescent readout of a C. difficile-specific protease, PPEP-1. NanoLuc, a bright and stable luciferase, was “caged” with a PPEP-1-responsive peptide tail that inhibited luminescence. Upon proteolytic cleavage, the peptide was released and NanoLuc activity was restored, providing a visible readout. The bioluminescent sensor detected PPEP-1 concentrations as low as 10 nM. Sensor uncaging was achieved within minutes, and signal was captured using a digital camera. Importantly, the sensor was also functional at ambient temperature and compatible with fecal material, suggesting that it can be readily deployed in a variety of settings.
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