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Methods and systems for detecting a pathogen in a biological sample

  • US 10,640,807 B2
  • Filed: 12/27/2012
  • Issued: 05/05/2020
  • Est. Priority Date: 12/29/2011
  • Status: Active Grant
First Claim
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1. A method for use with a blood sample, the method comprising:

  • staining the blood sample with at least Hoechst stain and Acridine Orange stain;

    using a microscope, acquiring a plurality of images of the blood sample, at least a first one of the images being acquired while the blood sample is illuminated by a light source having an illumination wavelength in which the Hoechst stain is visible, and at least a second one of the images being acquired while the blood sample is illuminated by a light source having an illumination wavelength in which the Acridine Orange stain is visible; and

    using at least one computer processor, detecting that a parasite is within a red blood cell within the blood sample, by;

    identifying an entity as a candidate of a parasite disposed within a red blood cell, by;

    identifying an object within the plurality of images as being a red blood cell candidate, based upon the object having characteristics of a red blood cell;

    identifying that a first stained area, which is stained by the Hoechst stain, is disposed within the red blood cell candidate, by analyzing the first one of the images; and

    identifying that a second stained area, which is stained by the Acridine Orange stain, is disposed within the red blood cell candidate, by analyzing the second one of the images; and

    validating that the entity is a parasite disposed within a red blood cell, by determining that structural features of the first and second stained areas satisfy predetermined criteria associated with the parasite, the structural features selected from the group consisting of;

    (i) sizes of the first stained area and the second stained area, and (ii) locations of the first stained area and the second stained area with respect to each other,the validating comprising distinguishing the entity from other entities within the blood sample that are stained by at least one of the Hoechst stain and the Acridine Orange.

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