Kits, compositions and methods for detecting a biological condition

Abstract

The present invention provides kits, apparatus and methods for determining a biological condition in a mammalian subject, the method includes incubating a specimen from a patient with at least one composition in a kit for a predetermined period of time to form at least one reaction product, when the subject has said biological condition, and receiving an indication of the at least one reaction product responsive to at least one reporter element in the kit thereby providing the indication of the biological condition in the subject.

1 - 13 . (canceled) 
     
     
         14 . A method for determining a biological condition in a subject, the method comprising:
 a. incubating a specimen from said subject in the kit of claim  1  for a predetermined period of time; and   b. receiving an indication responsive to said at least one reporter element thereby providing the indication of the biological condition in said subject.   
     
     
         15 . A method according to  claim 14 , wherein the biological condition is selected from blood diseases such as leukemia, thrombocytopenia, immune system disorders, local infections, urinary tract disorders, autoimmune diseases and sepsis. 
     
     
         16 . A method according to  claim 14 , wherein said indication is quantitative. 
     
     
         17 . A method according to  claim 14 , wherein the sample is of a volume of less than 200 microliters (μL). 
     
     
         18 . A method according to  claim 14 , wherein the method is completed within twenty minutes. 
     
     
         19 . A method according to  claim 18 , wherein the method is completed within fifteen minutes. 
     
     
         20 . A method for determining a biological condition in a mammalian subject, the method comprising:
 a. incubating a specimen from the subject with at least one composition in a kit for a predetermined period of time to form at least one reaction product, when said subject has said biological condition; and   b. receiving an indication of said at least one reaction product responsive to at least one reporter element in said kit thereby providing the indication of the biological condition in said subject.   
     
     
         21 . An automated method of determining the presence or absence of sepsis in a subject, comprising:
 a. contacting a blood sample from the subject with a fluorescently-labeled binding moiety specific to a sepsis marker, wherein the volume of the blood sample is 50 μL or smaller;   b. detecting the presence, absence or level of the binding moiety in the sample, thereby determining the presence or absence of sepsis in the subject within twenty minutes.   
     
     
         22 . The method of  claim 21 , wherein the sepsis marker is CD64. 
     
     
         23 . The method of  claim 21 , wherein the sepsis marker is CD163. 
     
     
         24 . The method of  claim 21 , further comprising contacting the blood sample with a second fluorescently-labeled binding moiety specific for a second sepsis marker. 
     
     
         25 . The method of  claim 24 , wherein the sepsis marker is CD64 and the second sepsis marker is CD163. 
     
     
         26 . The method of  claim 21 , wherein the binding moiety is an antibody. 
     
     
         27 . The method of  claim 21 , wherein the detecting step is performed in a device capable of receiving the sample and capable of detecting the binding moiety. 
     
     
         28 . The method of  claim 24 , further comprising the step of calibrating the device by detecting a population of the fluorescently-labeled particles. 
     
     
         29 . The method of  claim 28 , wherein said particles comprise the same fluorescent label as the fluorescently-labeled binding moiety. 
     
     
         30 . The method of  claim 29 , further comprising a second population of particles that comprise the same fluorescent label as the second fluorescently-labeled binding moiety. 
     
     
         31 . The method of  claim 28 , further comprising performing an internal calibration after said detecting said fluorescently-labeled binding moiety. 
     
     
         32 . The method of  claim 31 , wherein the calibration is completed in less than 5 minutes. 
     
     
         33 . The method of  claim 32 , wherein the particles are microbeads. 
     
     
         34 . The method of  claim 21 , wherein said method is performed in less than 15 minutes. 
     
     
         35 . The method of  claim 31 , wherein the particles are microbeads. 
     
     
         36 . The method of  claim 21 , further comprising the step of determining the presence of at least one cell population in the sample that is not bound by the binding moiety, thus providing an internal negative control for the sample. 
     
     
         37 . A disposable kit for rapid evaluation of a biological condition in a subject, the kit comprising:
 a static disposable cartridge adapted to receive a biological specimen from said subject, said cartridge including
 at least one treatment composition chamber adapted to house at least one composition, 
 a treatment chamber comprising at least one of a vortexing element and tortuous mixing means adapted to provide a rapid reaction of said specimen, said at least one composition to form a reaction product, when said subject has said biological condition, 
 a detection chamber adapted to receive said reaction product to provide an indication of said reaction product indicative of said biological condition, and 
 a plurality of valveless microfluidic channels, each adapted to convey at least one of said at least one treatment composition, said reaction product, at least one detector moiety and said reporter element between any two of said chambers, 
   wherein said chambers, elements and channels are activated according to a predefined sequence.   
     
     
         38 . A kit according to  claim 37 , further comprising instructions for obtaining an indication of the biological condition by using the kit. 
     
     
         39 . A kit according to  claim 37 , wherein said disposable cartridge comprises at least one of the following elements:
 a reservoir;   a pump;   a conduit;   a miniaturized flow cell;   a transport channel;   a reading channel;   a microfluidic element;   a compressed gas holding element;   a compressed gas releasing element;   a nozzle element; and   a mixing element.   
     
     
         40 . A kit according to  claim 39 , wherein said disposable microfluidics cartridge comprises at least two of the elements. 
     
     
         41 . A kit according to  claim 40 , wherein said disposable microfluidics cartridge comprises at least three of the elements. 
     
     
         42 . A kit according to  claim 37 , wherein said at least one composition disposed in said cartridge comprises at least one of:
 at least one target antibody;   at least one positive control identifying antibody; and   at least one negative control identifying detection moiety.   
     
     
         43 . A kit according to  claim 37 , wherein said at least one composition disposed in said cartridge comprises at least one reference composition comprising at least one of:
 a target signal reference composition; and   a reference identifier composition.   
     
     
         44 . A kit according to  claim 37 , wherein said at least one composition disposed in said cartridge comprises:
 a positive control moiety; and   a negative control moiety.   
     
     
         45 . A kit according to  claim 37 , wherein said at least one composition comprises a sepsis biomarker. 
     
     
         46 . A kit according to  claim 45 , wherein said biomarker comprises at least one of CD64 and CD163. 
     
     
         47 . A kit according to  claim 37 , wherein said predefined sequence is adapted to occur within fifteen minutes. 
     
     
         48 . A kit according to  claim 47 , wherein said rapid evaluation is adapted to occur within fifteen minutes. 
     
     
         49 . A kit according to  claim 37 , wherein the biological condition is selected from blood diseases such as leukemia, thrombocytopenia, immune system disorders, local infections, urinary tract disorders, autoimmune diseases and sepsis. 
     
     
         50 . A kit according to  claim 49 , wherein the biological condition is sepsis. 
     
     
         51 . A kit according to  claim 37 , wherein said indication is a visual indication. 
     
     
         52 . A kit according to  claim 50 , wherein said visual indication is indicative of said biological condition. 
     
     
         53 . A kit according to  claim 51 , wherein said visual indication is provided within fifteen minutes. 
     
     
         54 . A kit according to  claim 37 , wherein the sample of said biological specimen is of a volume of less than 200 microliters (μL). 
     
     
         55 . A kit according to  claim 37 , wherein said at least one composition is of a volume of less than 200 microliters (μL).