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.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4545073PMC
http://dx.doi.org/10.1063/1.4928879DOI Listing

Publication Analysis

Top Keywords

expressing blac
12
quantitative detection
8
detection cells
8
cells expressing
8
diagnosis tuberculosis
8
blac
5
blac droplet-based
4
droplet-based microfluidics
4
microfluidics diagnosis
4
tuberculosis paper
4

Similar Publications

Atypical Mycobacterium abscessus BlaRI Ortholog Mediates Regulation of Energy Metabolism but Not β-Lactam Resistance.

Mol Microbiol

October 2024

Division of Immunity and Pathogenesis, Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, Florida, USA.

Mycobacterium abscessus (Mab) is highly drug resistant, and understanding regulation of antibiotic resistance is critical to future antibiotic development. Regulatory mechanisms controlling Mab's β-lactamase (Bla) that mediates β-lactam resistance remain unknown. S.

View Article and Find Full Text PDF

Role of amygdala astrocytes in different phases of contextual fear memory.

Behav Brain Res

June 2024

Instituto de Farmacología Experimental de Córdoba (IFEC-CONICET) - Departamento de Farmacología Otto Orsingher (FCQ-UNC), Córdoba, Argentina. Electronic address:

Growing evidence indicates a critical role of astrocytes in learning and memory. However, little is known about the role of basolateral amygdala complex (BLA-C) astrocytes in contextual fear conditioning (CFC), a paradigm relevant to understand and generate treatments for fear- and anxiety-related disorders. To get insights on the involvement of BLA-C astrocytes in fear memory, fluorocitrate (FLC), a reversible astroglial metabolic inhibitor, was applied at critical moments of the memory processing in order to target the acquisition, consolidation, retrieval and reconsolidation process of the fear memory.

View Article and Find Full Text PDF

Objective: The emergence of immunotherapy has heralded a profound transformation in the therapeutic landscape of bladder cancer (BLAC). Immunotherapy, with its unique potential for "combination therapy", has brought about greater possibilities for treating BLCA. However, there is significant heterogeneity among bladder cancer patients, and a portion of those in advanced stages may not experience substantial benefits from chemotherapy.

View Article and Find Full Text PDF

High blood sugar and insulin insensitivity causes the lifelong chronic metabolic disease called Type 2 diabetes (T2D) which has a higher chance of developing different malignancies. T2D with comorbidities like Cancers can make normal medications for those disorders more difficult. There may be a significant correlation between comorbidities and have an impact on one another's health.

View Article and Find Full Text PDF

High-throughput fluorescent screening of β-lactamase inhibitors to improve antibiotic treatment strategies for tuberculosis.

Biosens Bioelectron

November 2022

Second Affiliated Hospital, Dalian Medical University, Dalian, 116023, China; School of Chemistry and Chemical Engineering, Henan Normal University, Xinxiang, 453007, China. Electronic address:

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb), which is a major threat to global public health. Currently, β-lactam antibiotics are rarely used in the treatment of TB, since Mtb naturally expresses β-lactamase (Blac) which renders Mtb resistant to such antibiotics due to β-lactam cleavage. Fortunately, antibiotic resistance can be overcome when β-lactam antibiotics are combined with a Blac inhibitor.

View Article and Find Full Text PDF

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!