Methods for improving assays of biological samples

• US 10,422,806 B1
• Filed: 07/25/2014
• Issued: 09/24/2019
• Est. Priority Date: 07/25/2013
• Status: Active Grant

• Current Assignee: Labrador Diagnostics LLC (SoftBank Group Corp.)
• Original Assignee: Theranos IP Company, LLC
• Inventors: Patel, Paul, Gibbons, Ian, Holmes, Elizabeth A.
• Primary Examiner(s): Cheu, Changwa J
• Application Number:
  US14/341,422
• Time in Patent Office: 1,887 Days
• Field of Search: None
• CPC Class Codes: G01N 2035/0094 optimisation; experiment design
• CPC Class Codes: G01N 33/56972 White blood cells
• CPC Class Codes: G01N 33/82 involving vitamins or their receptors
• CPC Class Codes: G01N 35/0092 Scheduling

Abstract

Methods for improving assays of biological samples are provided, including assays of small volume biological samples, such as blood samples obtained from a fingerstick. The methods include steps of selecting a baseline assay; providing a first revised assay by altering an aspect, a reagent, or a step of the baseline assay; comparing the results of the baseline assay and first revised assay; and identifying the first revised assay as an improved assay if the results of the first revised assay are similar to, are substantially the same as, or are better than, the results of the baseline assay. The methods may be iterated. Alterations include reducing sample volume; setting temperature; reducing step number and duration; altering reagent composition and number; and altering detection. Improved assays may reduce the duration, cost, or complexity of the assay, may improve assay sensitivity, accuracy, or reliability, and may provide synergistic improvements.

Claims

1. A method for improving an assay performed by an automatic assay device, said method comprising:

altering a baseline assay to provide a revised assay performed by an automatic assay device, said baseline assay having at least two assay steps and comprising an assay for detecting the presence or amount of an analyte in a biological sample having a sample volume,

said altering the baseline assay comprises;

reducing the number of assay steps, wherein said reducing the number of assay steps comprises combining two or more assay steps of said baseline assay by reducing the number of reagents used in the performance of the assay to provide that no more than two reagent mixing steps are required for use in said revised assay, said reducing the number of reagents comprises a) combining reagents of the baseline assay to provide a single combined reagent, or b) eliminating a reagent of the baseline assay or a component thereof; and

;

setting said sample volume to about 250 μ

L or less;

then

altering the dilution of said biological sample to provide a sample dilution of at least 10-fold in the revised assay;

then

setting the assay temperature of the revised assay to between about 32°

C. and about 37°

C.;

then

altering an assay step, altering the sequence of performance of assay steps, or both, of the baseline assay;

reducing the duration of an assay step effective that the time required to perform the revised assay is reduced as compared to the time required to perform the baseline assay;

then

requiring that said detection step comprises detecting an optical signal using an optical detector;

performing said revised assay on at least an aliquot of said diluted biological sample using an automatic assay device;

then

detecting results of the revised assay, wherein said detecting comprises detecting an optical signal;

then

comparing the results of said revised assay with the results of said baseline assay, and then

determining that the baseline assay is improved when the results of the revised assay are as good as, or better than, the baseline results,

wherein the revised assay comprises an improved assay when the baseline assay has been improved.

2. The method of claim 1, wherein detecting an optical signal comprises using an optical detector selected from a photomultiplier tube (PMT), a photodiode, a charge-coupled device (CCD), a photon counting detector, a camera, a microscope, and arrays and combinations thereof.

3. The method of claim 1, wherein the time required to perform said improved assay is less than about 30 minutes.

4. The method of claim 1, wherein performing the improved assay comprises three or fewer assay steps.

5. The method of claim 1, wherein the cost of the reagents required for performing the improved assay is less than the cost of reagents required for performing the baseline assay.

6. The method of claim 1, wherein the cost of the reagents required for performing the improved assay comprises about 1¢

or less per assay.

7. The method of claim 1, further comprising performing a second iteration of the steps of claim 1, wherein said improved assay obtained by the performance of the first iteration of the method is used as the baseline assay for the performance of the second iteration of the method.

8. A method for improving a baseline assay for detecting the presence or amount of a first analyte, said detecting being performed by an automatic assay device concurrently with assays for other analytes performed by said automatic assay device, said method comprising the steps of:

selecting a baseline assay having at least two assay steps; and

providing an improved assay based on said baseline assay, wherein said providing said improved assay comprises;

reducing the volume of the biological sample if the baseline assay sample volume is greater than about 250 μ

L;

then

eliminating pretreatment of said sample if the baseline assay includes a sample pretreatment step;

then

increasing the dilution of the biological sample by at least 10-fold to provide a diluted sample for providing an aliquot of diluted sample for use in the performance of said improved assay;

then;

eliminating a reagent, or combining two reagents to provide a single combined reagent, or a combination thereof, to provide that no more than two reagent mixing steps are required for the performance of said improved assay;

then

eliminating an assay step or combining two assay steps, or both, if the baseline assay includes more than three steps;

then

altering the assay temperature to be greater than or equal to 32°

C. if the baseline assay temperature is less than 32°

C.;

and then

reducing the time required to perform the assay to 30 minutes or less if the time required to perform the baseline assay time is greater than 30 minutes;

comparing the results of said improved assay with the results of said baseline assay, and then determining that the baseline assay is improved when the results of the improved assay are as good as, or better than, the baseline results;

thereby providing an improved assay for said first analyte, said improved assay being for concurrent use in an automatic assay device with other assays for other analytes in the same biological sample.

9. The method of claim 8, wherein said improved assay comprises an assay that works better in conjunction with other assays when said improved assay and said other assays are all performed concurrently using the same device or system to perform each of said assays using portions of the same biological sample.