Activated alkyne-enabled turn-on click bioconjugation with cascade signal amplification for ultrafast and high-throughput antibiotic screening.
Activated alkyne-enabled turn-on click bioconjugation with cascade signal amplification for ultrafast and high-throughput antibiotic screening.
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DOI：
10.1073/pnas.2302367120
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发表时间：
2023-07-04
影响因子：
11.1
通讯作者：
Zhu, Chunlei
中科院分区：
文献类型：
--
作者：
Lv, Shuyi;Wang, Chao;Xue, Ke;Wang, Jiaxin;Xiao, Minghui;Sun, Zhencheng;Han, Lei;Shi, Linqi;Zhu, Chunlei
关键词：
为了加快抗生素的发现，前提是建立一个通用的，敏感的，节省时间的方法进行抗菌药物敏感性测试。传统的基于荧光的细菌检测方法将整个细菌细胞视为一个不可分割的单元，并且细菌体内的大量生物底物未被利用。在此，我们提出了一种级联信号放大策略，用于简单，灵敏，超快和高通量的抗生素筛选，这是通过结合内部建立的用于分析物扩增的细菌裂解方法和新开发的用于荧光开启点击生物缀合的水溶性探针来实现的。该方法的检测限低至0.6 ~ 4 × 103 CFU/mL，整个操作过程不到3 h，在实际抗生素发现中具有很大的潜力。抗菌药物敏感性测试在发现新抗生素方面起着关键作用。然而，开发简单、敏感和快速的评估方法仍然具有挑战性。在此，我们报告了一种基于活化炔的级联信号放大策略，用于超快和高通量抗生素筛选。首先，合成了一种新型的水溶性聚集诱导发射（AIE）发光体，其含有活化的炔基，以使荧光开启和无金属点击生物缀合在生理条件下。利用内部建立的用于细菌裂解的方法，许多可点击的生物物质（即，细菌溶质和碎片）从细菌体中释放出来，这显著增加了分析物的量。通过活化的炔介导的开启点击生物缀合，由于增加的标记位点以及AIE效应，系统荧光信号被显著放大。这种级联信号放大策略有效地提高了检测灵敏度，从而能够进行超快的抗菌药物敏感性评估。通过与酶标仪集成，该方法进一步应用于高通量抗生素筛选。
To accelerate antibiotic discovery, the prerequisite is to establish a general, sensitive, and time-saving approach for antimicrobial susceptibility testing. Conventional fluorescence-based bacterial detection methods treat the entire bacterial cell as an indivisible unit, and the large quantities of biological substrates inside the bacterial bodies are unexploited. Herein, we present a cascade signal amplification strategy for simple, sensitive, ultrafast, and high-throughput antibiotic screening, which is achieved by combining the in-house established bacterial lysis method for analyte amplification and a newly developed water-soluble probe for fluorescence turn-on click bioconjugation. The determined limit of detection is as low as 0.6 to 4 × 103 CFU/mL, and the entire operation process is less than 3 h, holding great potential in real-world antibiotic discovery. Antimicrobial susceptibility testing plays a pivotal role in the discovery of new antibiotics. However, the development of simple, sensitive, and rapid assessment approaches remains challenging. Herein, we report an activated alkyne-based cascade signal amplification strategy for ultrafast and high-throughput antibiotic screening. First of all, a novel water-soluble aggregation-induced emission (AIE) luminogen is synthesized, which contains an activated alkyne group to enable fluorescence turn-on and metal-free click bioconjugation under physiological conditions. Taking advantage of the in-house established method for bacterial lysis, a number of clickable biological substances (i.e., bacterial solutes and debris) are released from the bacterial bodies, which remarkably increases the quantity of analytes. By means of the activated alkyne-mediated turn-on click bioconjugation, the system fluorescence signal is significantly amplified due to the increased labeling sites as well as the AIE effect. Such a cascade signal amplification strategy efficiently improves the detection sensitivity and thus enables ultrafast antimicrobial susceptibility assessment. By integration with a microplate reader, this approach is further applied to high-throughput antibiotic screening.
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影响因子：
3.1
作者：
Lee, Hwan Young;Park, Myung Jin;Shin, Kyoung-Jin
通讯作者：
Shin, Kyoung-Jin
影响因子：
16.6
作者：
Borkowski, Olivier;Koch, Mathilde;Faulon, Jean-Loup
通讯作者：
Faulon, Jean-Loup
影响因子：
6.7
作者：
Dhumal, Dinesh;Maron, Bar;Peng, Ling
通讯作者：
Peng, Ling
影响因子：
13.3
作者：
Bai, Haotian;Liu, Haixiang;Tang, Ben Zhong
通讯作者：
Tang, Ben Zhong
DOI：
10.3390/antibiotics10080974
发表时间：
2021-08-12
期刊：
Antibiotics (Basel, Switzerland)
影响因子：
--
作者：
Costa P;Gomes ATPC;Braz M;Pereira C;Almeida A
通讯作者：
Almeida A