Method for detecting infectious agents using computer controlled automated image analysis

Abstract

Computer controlled methods, systems and apparatus for detecting an infectious agent in a host cell, an animal cell, or plant cell, a body fluid or tissue sample to provide information useful to diagnose a disease or prognosticate disease susceptibility, extent or outcome are provided. In one illustrative embodiment, the infected animal cell is a Chlamydia infected mononuclear phagocyte in a blood sample. In another illustrative embodiment, the infected body fluid is a blood sample infected with “fee floating” Chlamydia . Detection and preferably, quantification of such an infected mononuclear phagocyte(s) or blood sample provides information useful in diagnosing or prognosticating susceptibility, extent or outcome of patients suspected of suffering from vascular, coronary or central nervous disease(s) or disorder(s).

1 . A computer software product, comprising a computer-readable storage medium having fixed therein a sequence of instructions which, when executed by a computer, direct the performance of a method which comprises: 
 a) acquiring image data of a sample of cells or a body fluid;    b) processing the image data to select and record images of a detectable signal indicative of an infectious agent of interest, if present;    c) quantifying the relative abundance of said infections agent if present; and    d) employing the relative abundance of the infectious agent, in combination with information relating to the physical or physiological state of a patient from whom the sample of cells or body fluid was obtained to determine a prognosis of susceptibility or diagnosis of a disease or disorder associated with the presence of said infectious agent.    
     
     
         2 . The computer software product of  claim 1 , wherein the infectious agent is a bacterium, mycoplasma, richkettsia, a spirochete, a fungus, a protozoal parasite, a virus or a prion.  
     
     
         3 . The computer software product of  claim 2 , wherein the infectious agent is  Chlamydia  sp. and the disorder is a vascular disorder.  
     
     
         4 . The computer software product of  claim 2 , wherein the infectious agent is  Borrelia  sp.  
     
     
         5 . A computer software product, comprising a computer-readable storage medium having fixed therein a sequence of instructions which, when executed by a computer, direct the performance of a method which comprises: 
 a) acquiring a microscope image of an optical field of a substrate having fixed thereon a monolayer of animal cells which either produce or are treated to produce a first signal specific to an animal cell of interest and treated to produce a second signal specific to an infectious agent of interest, if present, and transferring the image to an RGB image;    b) transferring the Red component of the RGB image to a new monochrome grey image;    c) transforming the grey level image to a binary image using a cut off point set to a value indicative of the expected size of the animal cells of interest;    d) operating on the binary image to remove noise and fill holes;    e) measuring size of the image of step d), selecting and recording areas representative of the animal cells of interest;    f) transferring the Red component of the original RGB image to a second binary image using an expected value indicative of the infectious agent of interest;    g) transferring the Green component of the original RGB image to grey level;    h) forming a new grey level image using all pixels having a value equal to a set value M and any grey level value in the Green component in the original RGB image equal to a set value N;    i) transforming the new grey level images to a third binary image;    j) operating on the third binary image to remove noise and fill holes;    k) recording the area of the third binary image after step j); and    l) determining whether or not, each identified animal cell is occupied by an infectious agent of interest.    
     
     
         6 . The computer software product of  claim 5 , wherein the method further comprises calculating the extent of each animal cell occupied by said infectious agent.  
     
     
         7 . The computer software product according to  claim 1 , further comprising a computer-readable storage medium having fixed therein a sequence of instructions which, when executed by a computer, detect or detect and quantify an infectious agent in an animal cell using a method as shown in  FIG. 2 .  
     
     
         8 . The computer software product according to  claim 1 , further comprising a computer-readable storage medium having fixed therein a sequence of instructions which, when executed by a computer, detect or detect and quantify an infectious agent in an animal cell using a method as shown in  FIG. 3 .  
     
     
         9 . The computer software product according to  claim 1 , further comprising a computer-readable storage medium having fixed therein a sequence of instructions which, when executed by a computer, detect or detect and quantify an infectious agent using a method as shown in  FIG. 4 .  
     
     
         10 . The computer software product according to  claim 5 , further comprising a computer-readable storage medium having fixed therein a sequence of instructions which, when executed by a computer, direct the performance of a method which comprises: 
 a) acquiring a microscope image of an optical field of a substrate having fixed thereon a sample of a body fluid or tissue from an animal patient suspected of containing an infectious agent of interest, which sample either produces or is treated to produce a first signal specific to an infectious agent of interest, if present, and transferring the image to an RGB image;    b) transferring the RGB image to an HLS image and transforming the HLS image to a new monochrome grey level image;    c) transforming the grey level image to a binary image;    d) operating on the binary image using at least one filter to identify blobs; and    e) selecting blobs based on the expected size of area of an infectious agent to determining whether or not an infectious agent of interest is present in the sample.    
     
     
         11 . A method for detecting an infectious agent in a sample of cells or a body fluid, comprising: a) acquiring a microscope image of an optical field of a substrate having fixed thereon a monolayer of animal cells which either produce or are treated to produce a first signal specific to an animal cell of interest and treated to produce a second signal specific to an infectious agent of interest, if present, and transferring the image to an RGB image; 
 b) transferring the Red component of the RGB image to a new monochrome grey image;    c) transforming the grey level image to a binary image using a cut off point set to a value indicative of the expected size of the animal cells of interest;    d) operating on the binary image to remove noise and fill holes;    e) measuring size of the image of step d),    f) selecting and recording areas representative of the animal cells of interest; transferring the Red component of the original RGB image to a second binary image using an expected value indicative of the infectious agent of interest;    g) transferring the Green component of the original RGB image to grey level;    h) forming a new grey level image using all pixels having a value equal to a set value M and any grey level value in the Green component in the original RGB image equal to a set value N;    i) transforming the new grey level images to a third binary image;    j) operating on the third binary image to remove noise and fill holes;    k) recording the area of the third binary image after step j); and    l) determining whether or not, each identified animal cell is occupied by an infectious agent of interest.    
     
     
         12 . The method of  claim 11 , which further comprises calculating the extent of each animal cell occupied by said infectious agent.  
     
     
         13 . The method according to  claim 11 , further comprising: 
 a) acquiring a microscope image of an optical field of a substrate having fixed thereon a sample of a body fluid or tissue from an animal patient suspected of containing an infectious agent of interest, which sample either produces or is treated to produce a first signal specific to an infectious agent of interest, if present, and transferring the image to an RGB image;    b) transferring the RGB image to an HLS image and transforming the HLS image to a new monochrome grey level image;    c) transforming the grey level image to a binary image;    d) operating on the binary image using at least one filter to identify blobs; and    e) selecting blobs based on the expected size of area of an infectious agent to determining whether or not an infectious agent of interest is present in the sample.    
     
     
         14 . The method according to  claim 11 , further comprising a method using the computer controlled method as shown in  FIG. 4 .  
     
     
         15 . The method according to  claim 11 , further comprising a method using the computer controlled method as shown in  FIG. 2 .  
     
     
         16 . The method according to  claim 11 , further comprising a method using the computer controlled method as shown in  FIG. 3 .