Quantitative detection of cells expressing BlaC using droplet-based microfluidics for use in the diagnosis of tuberculosis

使用基于液滴的微流体技术定量检测表达 BlaC 的细胞以用于结核病的诊断

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作者:Fengjiao Lyu, Manqi Xu, Yunfeng Cheng, Jinghang Xie, Jianghong Rao, Sindy K Y Tang

Abstract

This paper describes a method for the quantitative detection of cells expressing BlaC, a β-lactamase naturally expressed by Mycobacterium tuberculosis, intended for the diagnosis of tuberculosis. The method is based on the compartmentalization of bacteria in picoliter droplets at limiting dilutions such that each drop contains one or no cells. The co-encapsulation of a fluorogenic substrate probe for BlaC allows the quantification of bacteria by enumerating the number of fluorescent drops. Quantification of 10 colony forming units per milliliter is demonstrated. Furthermore, the encapsulation of single cell in drops maintains the specificity of the detection scheme even when the concentration of bacteria that do not express BlaC exceeds that expressing BlaC by one million-fold.

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