Process control for increased robustness of genetic assays

• US 10,429,399 B2
• Filed: 09/18/2015
• Issued: 10/01/2019
• Est. Priority Date: 09/24/2014
• Status: Active Grant

• Current Assignee: Invitae Corporation
• Original Assignee: Good Start Genetics, Inc. (Invitae Corporation)
• Inventors: Boyden, Eric D.
• Primary Examiner(s): Visone, Thomas J.
• Application Number:
  US14/858,637
• Publication Number:
  US 20160084854A1
• Time in Patent Office: 1,474 Days
• Field of Search: None
• CPC Class Codes: C12N 15/1003 Extracting or separating nucleic acids from biological samples, e.g. pure separation or isolation methods; Conditions, buffers or apparatuses therefor
• CPC Class Codes: C12Q 1/6806 Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay C12Q1/6804 takes precedence
• CPC Class Codes: G01N 1/30 Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
• CPC Class Codes: G01N 21/80 Indicating pH value
• CPC Class Codes: G01N 31/221 for investigating pH value
• CPC Class Codes: G01N 33/689 related to pregnancy or the gonads
• CPC Class Codes: G01N 33/84 involving inorganic compounds or pH

Abstract

The present disclosure describes a method of processing a biological sample. The method includes contacting the sample with a pH indicator that changes color in response to changes in pH. The presence of a pH indicator such as cresol red provides the benefit of improved visualization of the sample, verification that the sample is properly frozen and thawed, and increased precision in pH adjustments during the assay, among other benefits.

Claims

1. A method for processing a biological sample, the method comprising:

obtaining a sample comprising at least one cell biopsied from an embryo in a solution of 2-3 μ

l of liquid;

contacting the sample with an indicator dye comprising cresol red that provides a visual indication that the sample is frozen and within a specific, pre-defined pH range;

exposing the sample to a freezing temperature;

determining based on the visual indication that the sample is frozen; and

performing a preimplantation genetic screening procedure comprising;

using a camera controlled by computer software to evaluate a color of the sample;

confirming with the computer software that the color indicates that the sample remains frozen;

contacting the sample with an alkaline lysis solution followed by contacting the sample with an acid to achieve a neutral pH;

using the camera and the computer software to evaluate a color of the sample and confirm that the sample has a neutral pH; and

analyzing a karyotype of the at least one biopsied cell in the sample.

2. The method of claim 1, further comprising:

assessing said indicator in order to determine if the pH range is appropriate; and

delivering the sample to a laboratory if said indicator indicates that the sample is frozen.

3. The method of claim 1, further comprising the step of assessing said indicator in order to determine a position of said cells in a vial containing the sample.

4. The method of claim 1, wherein the alkaline lysis solution contains said indicator;

and wherein the acid contains said indicator.

5. The method of claim 2, wherein said assessing step is performed by observing a color change in the sample.

6. The method of claim 5, wherein color change is monitored by automatic means, wherein the automatic means comprises a camera.

7. The method of claim 2, wherein said assessing step is performed by monitoring electrical conductivity in the sample.