System and methods for bilirubin analysis

• US 10,324,097 B2
• Filed: 04/16/2018
• Issued: 06/18/2019
• Est. Priority Date: 08/30/2017
• Status: Active Grant

• Current Assignee: NeoLight, LLC
• Original Assignee: NeoLight, LLC
• Inventors: Palaniswamy, Sivakumar, Krishnaraju, Deepakshyam, Peacock, Martin
• Primary Examiner(s): Xu, Xiaoyun R
• Application Number:
  US15/954,360
• Publication Number:
  US 20190064190A1
• Time in Patent Office: 428 Days
• Field of Search: None
• CPC Class Codes: B01L 2200/04 Exchange or ejection of cartridges, containers or reservoirs
• CPC Class Codes: B01L 2200/147 Employing temperature sensors
• CPC Class Codes: B01L 2200/16 Reagents, handling or storing thereof
• CPC Class Codes: B01L 2300/021 Identification, e.g. bar codes
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/18 Means for temperature control
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• CPC Class Codes: G01N 27/308 at least partially made of carbon
• CPC Class Codes: G01N 27/3277 being a redox reaction, e.g. detection by cyclic voltammetry voltammetry per se G01N27/42, G01N27/48
• CPC Class Codes: G01N 27/48 using polarography, i.e. measuring changes in current under a slowly-varying voltage
• CPC Class Codes: G01N 2800/085 Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/49 Blood chemical methods for determining blood cell populations G01N33/5094; chemical analysis of blood groups or blood types G01N33/80
• CPC Class Codes: G01N 33/728 Bilirubin; including biliverdin

Abstract

A method for measuring bilirubin levels in a subject. The method can include the steps of providing a sample to be measured from the subject, wherein the sample comprises bilirubin bound to albumin; adding a release agent to the sample, the release agent configured to release the bound bilirubin from the albumin; measuring electrochemical data of the sample using an electrochemical cell; and determining a total serum bilirubin concentration of the sample using the electrochemical data. This method is capable of providing a simpler, faster, and more robust measurement when compared to traditional bilirubin assay methods.

Claims

1. A method for measuring bilirubin levels in a subject, the method comprising:

providing a fluid sample to be measured from the subject to a microfluidic device, wherein the sample comprises bilirubin bound to albumin;

contacting the fluid sample and a release agent in the form of a dry solid, the release agent configured to release the bound bilirubin from the albumin;

measuring electrochemical data of the sample using an electrochemical cell, wherein the electrochemical cell comprises screen printed electrodes; and

determining a total serum bilirubin concentration of the sample using the electrochemical data.

2. The method as recited in claim 1, wherein the fluid sample is an untreated blood sample of the subject.

3. The method as recited in claim 1, wherein the release agent is 4,4′

-(1,1-dioxido-3H-2,1-benzoxathiole-3,3-diyl)bis(2,6-dibromophenol).

4. The method as recited in claim 1, wherein the electrochemical data is voltammetric data.

5. The method as recited in claim 1, further comprising adding additional agents to the sample, wherein the addition of the agents occurs prior to the measurement of the electrochemical data.

6. The method as recited in claim 5, wherein the additional agents are in the form of dry solids.

7. The method as recited in claim 5, wherein the additional agents include at least one of a surfactant, buffer, salt, or second release agent.

19. The method of claim 1, wherein the volume of the sample is less than 40 microliters.

8. A method for diagnosing a subject based on electrochemical detection of analytes, the method comprising:

providing a blood sample to be measured from the subject to a microfluidic device, wherein the sample comprises both unbound bilirubin and bilirubin bound to albumin;

contacting the blood sample and a release agent in the form of a dry solid, the release agent configured to release the bound bilirubin from the albumin;

measuring electrochemical data of the sample using an electrochemical cell, wherein the electrochemical cell comprises screen printed electrodes;

determining concentrations of conjugated bilirubin, unconjugated bilirubin, total serum bilirubin, and albumin using the electrochemical data; and

making a diagnosis of an illness for the subject based on the determined concentrations of conjugated bilirubin, unconjugated bilirubin, total serum bilirubin, and albumin.

9. The method as recited in claim 8, wherein the blood sample is an untreated blood sample of the subject.

10. The method as recited in claim 8, wherein the release agent is 4,4′

-(1,1-dioxido-3H-2,1-benzoxathiole-3,3-diyl)bis(2,6-dibromophenol).

11. The method as recited in claim 8, wherein the electrochemical data is voltammetric data.

12. The method of claim 8, wherein the illness is icterus.

13. The method as recited in claim 8, further comprising adding additional agents to the sample, wherein the addition of the agents occurs prior to the measurement of the electrochemical data.

14. The method as recited in claim 13, wherein the additional agents are in the form of dry solids.

15. The method as recited in claim 13, wherein the additional agents include at least one of a surfactant, buffer, salt, or second release agent.

16. The method as recited in claim 8, wherein the temperature of the sample is actively controlled between 20 and 40 degrees Celsius while the step of measuring the electrochemical data is occurring.

17. The method as recited in claim 8, further comprising filtering the sample, wherein the filtering occurs prior to the measurement of the electrochemical data.

18. The method as recited in claim 8, further comprising providing a treatment to the subject, wherein at least one of the type of treatment and intensity of treatment is determined using the diagnosis of the illness.

20. The method of claim 8, wherein the volume of the blood sample is less than 40 microliters.