Systems and Methods for Detecting a Biological Condition

Abstract

The present invention provides self-contained systems, apparatus and methods for determining a chemical state, the system includes a stationary cartridge for performing the assay therein, the cartridge adapted to house at least one reagent adapted to react with a sample; and at least one reporter functionality adapted to report a reaction of the at least one reagent with the sample to report a result of the assay, a mechanical controller including a first urging means adapted to apply a force externally onto the cartridge to release the at least one reagent; and at least one second urging means adapted to apply a removable force to induce fluidic movement in a first direction in the cartridge and upon removal of the force causing fluidic movement in an opposite direction to the first direction, an optical reader adapted to detect the reaction and a processor adapted to receive data from the optical reader and to process the data to determine said chemical state.

1 . A flow cytometric assay method for determining a biological condition in a subject, the method comprising:
 a. incubating a sample from said subject in a self-contained stationary cartridge housing at least one composition and at least one reporter functionality, adapted to report a reaction of said at least one composition with said sample, with said at least one composition and with said at least one reporter functionality to report an assay of said reaction;   b. optically detecting said reaction in a moving fluid;   c. processing data outputted in said optically detecting step to determine said assay, comprising:
 i. processing a time series from each channel of an at least eight channel photomultiplier array or light receiving reader unit and scatter channels data; 
 ii. characterizing at least one event in terms of a fluor composition thereof to output at least one derived time series; and 
 iii. detecting at least one particle type from said at least one event associated with said at least one derived time series; and 
   d. receiving an indication responsive to said assay, thereby providing an indication of the biological condition in said subject.   
     
     
         2 . A method according to  claim 1 , wherein said at least one composition is wet. 
     
     
         3 . A method according to  claim 1 , wherein said at least one composition is dry. 
     
     
         4 . A method according to  claim 1 , wherein said at least one particle type is from said sample, said at least one particle type selected from the group consisting of: a neutrophil, a monocyte, a bead, a lymphocyte, a reject and combinations thereof. 
     
     
         5 . A method according to  claim 1 , wherein said sample is in a form selected from the group consisting of: a solid, a powder, a crystal form, a liquid, a colloid, a suspension and combinations thereof. 
     
     
         6 . A method according to  claim 1 , wherein said optically detecting step comprises:
 i. providing an excitation beam to said cartridge;   ii. measuring a forward scatter measurement from particles in at least one of said sample and said at least composition in an optical reader;   iii. passing a returned beam from said sample via a high-pass filter and a concave grating; and   iv. detecting a plurality of at least eight spectrally distinct signals associated with said at least one reporter functionality produced by said concave grating in said at least one of photomultiplier array (PMT) and a light-receiving reader unit and scatter channels data.   
     
     
         7 . A method according to  claim 1 , wherein said, wherein the biological condition is selected from blood diseases, leukemia, thrombocytopenia, immune system disorders, local infections, urinary tract disorders, autoimmune diseases and sepsis. 
     
     
         8 . A method according to  claim 1 , wherein said optically detecting step comprises detecting a signal associated with at least one reporter functionality, said at least one reporter functionality adapted to report a reaction of said at least one composition with said sample, herein said at least one composition comprises:
 i. a cell surface marker;   ii. a cell stain;   iii. a reagent bound to a solid support;   iv. a chemical indicator; and   v. a biological cell indicator;   
       wherein said reagent bound to said solid support is selected from the group consisting of an immobilized enzyme, an immobilized substrate, a plasma protein bead, an antibody bead and an antigen bead. 
     
     
         9 . A method according to  claim 8 , wherein said biological cell indicator is selected from the group consisting of a cell cycle stage indicator, a cell proliferation indicator, a cytokine indicator, a metabolic indicator and an apoptosis indicator. 
     
     
         10 . A method according to  claim 8 , wherein said at least one composition comprises at least two compositions. 
     
     
         11 . A method according to  claim 10 , wherein said at least two compositions comprise at least one of:
 a. a cell surface marker and a cell element stain;   b. a cell surface marker and a plasma protein bead assay;   c. a cell surface marker and a solution change marker;   d. a cell element stain and a plasma protein bead assay; and   e. a cell element stain and a solution change marker.   
     
     
         12 . A method according to  claim 8 , wherein said at least one composition comprises:
 a) a detection composition comprising at least one of;
 i. at least one target antibody; 
 ii. at least one positive control identifying antibody; and 
 iii. at least one negative control identifying detection moiety or characteristic; and 
   b) at least one reference composition comprising at least one of;
 i. a target signal reference composition; and 
 ii. a reference identifier composition. 
   
     
     
         13 . A method according to  claim 8 , wherein said at least one composition comprises:
 a) an antibody composition comprising at least one of;
 i. at least one target antibody; 
 ii. at least one positive control identifying antibody; and 
 iii. at least one negative control identifying antibody or characteristic; and 
   b) at least one reference composition comprising at least one of;
 iv. a target signal reference composition; and 
 v. a reference identifier composition. 
   
     
     
         14 . A method according to  claim 8 , wherein said sample comprises at least one of;
 i. a bodily specimen comprising a target moiety;   ii. a positive control moiety; and   iii. a negative control moiety.   
     
     
         15 . A method according to  claim 8 , wherein the at least one composition further comprises at least one conditioning moiety comprising;
 a. at least one lysis reagent; and   b. at least one diluent.   
     
     
         16 . A method according to  claim 6 , wherein said providing step further comprises passing said excitation beam via a dichroic mirror through an objective onto a reading channel in said cartridge. 
     
     
         17 . A method according to  claim 16 , further comprising, collecting particle fluorescent emission through said objective, further passing said particle fluorescent emission through said dichroic mirror and reflecting said fluorescent emission from a beamsplitter into a detection path. 
     
     
         18 . A method according to  claim 1 , wherein said receiving an indication step further comprises outputting bandwidth leveled and smoothed arrays. 
     
     
         19 . A method according to  claim 18 , wherein said receiving an indication step further comprises outputting a three-dimensional graph showing optical output over time from said optically detecting step. 
     
     
         20 . A method according to  claim 18 , wherein said receiving an indication step further comprises providing a graphical display showing a cluster analysis of wavebands from at least some of said eight channels. 
     
     
         21 . A method according to  claim 20 , further comprising applying an algorithm to said cluster analysis to provide a detection of at least one signature associated with said biological condition.