US 12,352,714 B2
Devices, systems and methods to detect the presence of ß-lactam antibiotic hydrolyzing bacteria in a sample
Meike Herget, Woodside, CA (US); Oren S. Knopfmacher, San Francisco, CA (US); and Michael D. Laufer, Menlo Park, CA (US)
Assigned to Avails Medical, Inc., Menlo Park, CA (US)
Filed by Avalis Medical, Inc., Menlo Park, CA (US)
Filed on Jun. 13, 2019, as Appl. No. 16/440,829.
Application 16/440,829 is a continuation of application No. PCT/US2016/066315, filed on Dec. 13, 2016.
Prior Publication US 2019/0310214 A1, Oct. 10, 2019
Int. Cl. G01N 27/02 (2006.01); C12N 1/06 (2006.01); C12Q 1/18 (2006.01); C12Q 1/34 (2006.01)
CPC G01N 27/021 (2013.01) [C12N 1/06 (2013.01); C12Q 1/18 (2013.01); C12Q 1/34 (2013.01)] 13 Claims
OG exemplary drawing
 
1. A method of detecting the presence of β-lactam antibiotic hydrolyzing bacteria in a sample, the method comprising:
introducing the sample comprising the β-lactam antibiotic hydrolyzing bacteria to a container housing a filter, wherein the filter comprises a filter surface, wherein the filter surface is a mesh or matrix structure made of a polymeric material, wherein the filter is secured by the container housing;
introducing a buffer solution to the β-lactam antibiotic hydrolyzing bacteria, wherein the buffer solution comprises a β-lactamase substrate, a β-lactamase activator, and a lysing agent to lyse cells of the bacteria to release-lactam antibiotic hydrolyzing enzymes,
wherein the buffer solution and the B-lactam antibiotic hydrolyzing enzymes form a reaction mixture;
exposing a sensor having an electrical characteristic to the reaction mixture, wherein the sensor comprises an active electrode and a quasi-reference electrode, wherein the active electrode and the quasi-reference electrode are disposed on a base dielectric layer; and
monitoring a change in the electrical characteristic of the sensor to detect the presence of the β-lactam antibiotic hydrolyzing bacteria in the sample originally introduced, wherein the change in the electrical characteristic is a change in a voltage potential between the active electrode and the quasi-reference electrode measured by a parameter analyzer coupled to the active electrode and the quasi-reference electrode.