The Texas A&m University System

Leland Stanford Junior University

Jeffrey D Cirillo

Jianghong Rao

Hexin Xie

James C Sacchettini

Yunfeng Cheng

Michael T Norman

The present disclosure provides methods, reagents, systems, and devices that target &#x3b2; lactamase as a biomarker for the sensitive and specific detection of tuberculosis-complex bacteria. Specifically, the present disclosure relates to methods and compositions for the detection of specific &#x3b2;-lactamase protein and nucleic acid sequences to indicate the presence of tuberculosis-complex bacteria.

Beta lactamase as biomarker for the specific detection of tuberculosis-complex bacteria

&#x3b2;-Lactamase substrates and methods for using the substrates to detect &#x3b2;-lactamase and to diagnose tuberculosis.

2,7-disubstituted cephalosporin derivatives as &#x3b2;-lactamase substrates and methods for their use for the diagnosis of tuberculosis

2,7-disubstituted cephalosporin derivatives as beta-lactamase substrates and methods for their use for the diagnosis of tuberculosis

&#x3b2;-Lactamase substrates and methods for using the substrates to detect &#x3b2;-lactamase diagnose tuberculosis.

&#x3b2;-lactamase substrates and methods of their use for the diagnosis of tuberculosis

Provided herein are assays and in vitro methods to determine susceptibility to a drug effective against a pathogenic bacteria, for example, a pathogenic Mycobacteria, that has a beta-lactamase activity. An excitation wavelength is delivered to a biological sample obtained from a subject having an infection from the pathogenic bacteria in the presence of a beta-lactamase substrate. The intensity of a signal, such as a fluorescent, luminescent or colorimetric signal, at an emission wavelength of a product of the beta-lactamase on the subject is correlated to drug susceptibility. Also provided is an assay system for monitoring drug susceptibility of a pathogenic bacteria comprising color-producing substrates for a beta-lactamase of the pathogenic bacteria, an assay device for visibly detecting a product of the beta-lactamase on the substrate thereof and a reader configured to quantify the visibly detected product.

Use of bacterial beta-lactamase for in vitro diagnostics and in vivo imaging, diagnostics and therapeutics

Provided herein are methods for detecting, quantifying, differentiating, diagnosing and imaging pathogenic bacteria or condition associated therewith using substrates for bacterial enzymes. Fluorescent, luminescent or colorimetric signals emitted by substrates or enzyme products in the presence of the bacteria are compared to controls to detect and locate the pathogenic bacteria. Provided is a method for screening therapeutic agents to treat the pathophysiological conditions by measuring a signal emitted from the substrates or products in the presence and absence of the potential therapeutic agent and a diagnostic method for detecting a mycobacterial infection in a subject by contacting biological samples with a substrate and imaging for signals emitted from a mycobacterial beta-lactamase product. Also provided are fluorogenic substrates or substrates comprising a colored dye or a chemical reagent effective to induce a color or pH change.

Growing community of inventors

College Station, TX, United States of America

Jeffrey D Cirillo