US2023227887A1PendingUtilityA1

Therapeutic Drug Monitoring

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Individually held — no corporate assignee on recordPriority: Jun 2, 2017Filed: Oct 31, 2022Published: Jul 20, 2023
Est. expiryJun 2, 2037(~10.8 yrs left)· nominal 20-yr term from priority
Inventors:Cidalia Irene Azevedo Pina VazAcacio Agostinho Goncalves Rodrigues
C12Q 1/18G01N 15/1456G01N 2015/1006G01N 33/582
50
PatentIndex Score
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Claims

Abstract

Disclosed herein are methods and compositions for antimicrobial quantification and functional measurement. In one aspect, a method for quantifying antimicrobial comprises: obtaining a biological sample from a patient receiving an antimicrobial; incubating the biological sample with a reference microbial strain and a fluorophore for detecting cell lesion; measuring a first signal of fluorescent intensity in the incubated biological sample using flow cytometry; and comparing the first signal to a calibrating curve previously generated for the antimicrobial, thereby quantifying the antimicrobial present in the biological sample.

Claims

exact text as granted — not AI-modified
1 . A method for quantifying an antimicrobial in a biological sample, comprising:
 incubating a biological sample with a reference microbial strain and a fluorophore for detecting cell lesion, wherein the biological sample is obtained from a patient receiving an antimicrobial;   measuring, using flow cytometry, a first signal of fluorescent intensity of the fluorophore in the biological sample incubated with the reference microbial strain; and   comparing the first signal to a calibrating curve previously generated for the antimicrobial against the reference microbial strain, thereby quantifying the antimicrobial present in the biological sample.   
     
     
         2 . The method of  claim 1 , wherein the biological sample is one or more of serum, plasma, urine, spinal fluid, bronchial secretions or washing or any other body fluid. 
     
     
         3 . The method of  claim 1 , wherein the patient has an infection. 
     
     
         4 . The method of  claim 1 , wherein the antimicrobial is an antibiotic or antifungal. 
     
     
         5 . The method of  claim 1 , wherein the reference microbial strain is  E .  coli  or  S .  aureus . 
     
     
         6 . The method of  claim 1 , wherein the fluorophore is selected from DiBAC 4 , DIOC, propidium iodide, DHR, C-FDA or FUN-1. 
     
     
         7 . The method of  claim 1 , wherein the incubating step comprises incubating for about 60 minutes. 
     
     
         8 . The method of  claim 1 , wherein the calibrating curve is generated using different concentrations of the antimicrobial incubated with the reference microbial strain. 
     
     
         9 . The method of  claim 1 , further comprising incubating a separate aliquot of the biological sample with a microbial strain isolated from the patient and the fluorophore, and measuring a second signal of fluorescent intensity in the separate aliquot using flow cytometry. 
     
     
         10 . The method of  claim 9 , wherein the calibrating curve is generated using different concentrations of the antimicrobial incubated with the microbial strain isolated from the patient. 
     
     
         11 . The method of  claim 9 , wherein the microbial strain isolated from the patient is E.  coli  or S.  aureus . 
     
     
         12 . A method for quantifying an antimicrobial in a biological sample, comprising:
 incubating a biological sample obtained from a patient receiving an antimicrobial with a reference microbial strain and a fluorophore for detecting a cell lesion;   before an effect on microbial growth becomes apparent, measuring a first signal of fluorescent intensity of the fluorophore in the biological sample incubated with the reference microbial strain using flow cytometry; and   comparing the first signal to a calibrating curve previously generated for the antimicrobial against the reference microbial strain, thereby quantifying the antimicrobial present in the biological sample.   
     
     
         13 . The method of  claim 12 , wherein the biological sample is one or more of serum, plasma, urine, spinal fluid, bronchial secretions or washing or any other body fluid. 
     
     
         14 . The method of  claim 12 , wherein the patient has an infection. 
     
     
         15 . The method of  claim 12 , wherein the antimicrobial is an antibiotic or antifungal. 
     
     
         16 . The method of  claim 12 , wherein the reference microbial strain is  E .  coli  or  S .  aureus . 
     
     
         17 . The method of  claim 12 , wherein the fluorophore is selected from DiBAC 4 , DIOC, propidium iodide, DHR, C-FDA or FUN-1. 
     
     
         18 . The method of  claim 12 , wherein the incubating step comprises incubating for about 60 minutes. 
     
     
         19 . The method of  claim 12 , wherein the calibrating curve is generated using different concentrations of the antimicrobial incubated with the reference microbial strain. 
     
     
         20 . The method of  claim 12 , further comprising incubating a separate aliquot of the biological sample with a microbial strain isolated from the patient and the fluorophore, and measuring a second signal of fluorescent intensity in the separate aliquot using flow cytometry. 
     
     
         21 . The method of  claim 20 , wherein the calibrating curve is generated using different concentrations of the antimicrobial incubated with the microbial strain isolated from the patient. 
     
     
         22 . The method of  claim 20 , wherein the microbial strain isolated from the patient is  E .  coli  or  S .  aureus .

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