Development of a Highly Specific, Selective, and Sensitive Fluorescent Probe for Detection of Mycobacteria in Human Tissues

开发一种用于检测人体组织中分枝杆菌的高特异性、选择性和灵敏性的荧光探针

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作者:Siddhi Gupta, Deepak Kumar Mishra, Mehak Zahoor Khan, Varsha Saini, Devashish Mehta, Sandeep Kumar, Aditya Yadav, Madhurima Mitra, Parul Rani, Mukesh Singh, Chayan Kanti Nandi, Prasenjit Das, Vineet Ahuja, Vinay Kumar Nandicoori, Avinash Bajaj

Abstract

Tuberculosis (TB), including extrapulmonary TB, is responsible for more than one million deaths in a year worldwide. Existing methods of mycobacteria detection have poor sensitivity, selectivity, and specificity, especially in human tissues. Herein, the synthesis of a cholic acid-derived fluorescent probe (P4) that can specifically stain the mycobacterium species is presented. It is shown that P4 probe specifically binds with mycobacterial lipids, trehalose monomycolate, and phosphatidylinositol mannoside 6. P4 probe can detect mycobacteria in polymicrobial planktonic cultures and biofilms with high specificity, selectivity, and sensitivity. Moreover, it can detect a single mycobacterium in the presence of 10 000 other bacilli. Unlike the probes that depend on active mycobacterial enzymes, the membrane-specific P4 probe can detect mycobacteria even in formalin-fixed paraffin-embedded mice and human tissue sections. Therefore, the ability of the P4 probe to detect mycobacteria in different biological milieu makes it a potential candidate for diagnostic and prognostic applications in clinical settings.

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