Hybrid automated continuous nucleic acid and protein analyzer using real-time PCR and liquid bead arrays

Abstract

A nucleic acid assay system for analyzing a sample using a reagent. A sample and reagent delivery unit is operatively connected to a thermal cycler for delivering the sample and the reagent to the thermal cycler. A hybridization chamber is operatively connected to the thermal cycler. A flow cytometer is operatively connected to the hybridization chamber.

1 . A nucleic acid assay apparatus for analyzing a sample using a reagent, comprising: 
 a thermal cycler,    a sample and reagent delivery unit operatively connected to said thermal cycler for delivering the sample and the reagent to said thermal cycler,    a hybridization chamber operatively connected to said thermal cycler, and    a flow cytometer operatively connected to said hybridization chamber.    
     
     
         2 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with each bead having a unique spectral address.  
     
     
         3 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize polystyrene latex microspheres beads with each bead having a unique spectral address.  
     
     
         4 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with varying ratios of red and orange infrared dyes giving each bead a unique spectral address.  
     
     
         5 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize a 100-plex array of beads with varying ratios of red and orange infrared dyes giving each bead a unique spectral address.  
     
     
         6 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with each bead having a capture antibody specific for a target antigen.  
     
     
         7 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with at least one bead having an anthrax capture antibody specific for a target anthrax antigen.  
     
     
         8 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with at least one bead having a plague capture antibody specific for a target plague antigen.  
     
     
         9 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with at least one bead having a small pox capture antibody specific for a target small pox antigen.  
     
     
         10 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with at least one bead having a botox capture antibody specific for a target botox antigen.  
     
     
         11 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with each bead having a capture antibody specific for a target antigen and a fluorescent reporter.  
     
     
         12 . The nucleic acid assay apparatus of  claim 1  wherein said flow cytometer utilizes at least one laser.  
     
     
         13 . The nucleic acid assay apparatus of  claim 1  wherein said flow cytometer utilizes a red laser.  
     
     
         14 . The nucleic acid assay apparatus of  claim 1  wherein said flow cytometer utilizes a green laser.  
     
     
         15 . The nucleic acid assay apparatus of  claim 1  wherein said flow cytometer utilizes a red laser and a green laser.  
     
     
         16 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with each bead having a fluorescent reporter and said flow cytometer utilizes at least one laser for bead interrogation by fluoresce of said fluorescent reporter.  
     
     
         17 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with each bead having a capture antibody specific for a target antigen and a fluorescent reporter and said flow cytometer utilizes at least one laser for bead interrogation.  
     
     
         18 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber and said flow cytometer utilize beads with each bead having a capture antibody specific for a target antigen and a fluorescent reporter and said flow cytometer utilizes a green laser for bead interrogation by fluoresce of said fluorescent reporter.  
     
     
         19 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber includes a heater.  
     
     
         20 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber includes a fan.  
     
     
         21 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber includes a heater and a fan.  
     
     
         22 . The nucleic acid assay apparatus of  claim 1  wherein said hybridization chamber includes a heater, a fan, and a temperature control sensor.  
     
     
         23 . A nucleic acid assay apparatus for analyzing a sample using a reagent, comprising: 
 thermal cycler means,    sample and reagent delivery means operatively connected to said thermal cycler means for delivering the sample and the reagent to said thermal cycler means,    hybridization means operatively connected to said thermal cycler means, and    flow cytometer means operatively connected to said hybridization means.    
     
     
         24 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with each bead having a unique spectral address.  
     
     
         25 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize polystyrene latex microspheres beads with each bead having a unique spectral address.  
     
     
         26 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with varying ratios of red and orange infrared dyes giving each bead a unique spectral address.  
     
     
         27 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize a 100-plex array of beads with varying ratios of red and orange infrared dyes giving each bead a unique spectral address.  
     
     
         28 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with each bead having a capture antibody specific for a target antigen.  
     
     
         29 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with at least one bead having an anthrax capture antibody specific for a target anthrax antigen.  
     
     
         30 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with at least one bead having a plague capture antibody specific for a target plague antigen.  
     
     
         31 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with at least one bead having a small pox capture antibody specific for a target small pox antigen.  
     
     
         32 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with at least one bead having a botox capture antibody specific for a target botox antigen.  
     
     
         33 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with each bead having a capture antibody specific for a target antigen and a fluorescent reporter.  
     
     
         34 . The nucleic acid assay apparatus of  claim 23  wherein said flow cytometer means utilizes at least one laser.  
     
     
         35 . The nucleic acid assay apparatus of  claim 23  wherein said flow cytometer means utilizes a red laser.  
     
     
         36 . The nucleic acid assay apparatus of  claim 23  wherein said flow cytometer means utilizes a green laser.  
     
     
         37 . The nucleic acid assay apparatus of  claim 23  wherein said flow cytometer means utilizes a red laser and a green laser.  
     
     
         38 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with each bead having a fluorescent reporter and said flow cytometer means utilizes at least one laser for bead interrogation by fluoresce of said fluorescent reporter.  
     
     
         39 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with each bead having a capture antibody specific for a target antigen and a fluorescent reporter and said flow cytometer means utilizes at least one laser for bead interrogation.  
     
     
         40 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means and said flow cytometer means utilize beads with each bead having a capture antibody specific for a target antigen and a fluorescent reporter and said flow cytometer means utilizes a green laser for bead interrogation by fluoresce of said fluorescent reporter.  
     
     
         41 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means includes a heater.  
     
     
         42 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means includes a fan.  
     
     
         43 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means includes a heater and a fan.  
     
     
         44 . The nucleic acid assay apparatus of  claim 23  wherein said hybridization means includes a heater, a fan, and a temperature control sensor.  
     
     
         45 . A nucleic acid assay method for analyzing a sample using a reagent, comprising the steps of: 
 providing a thermal cycler,    providing a hybridization unit,    providing a flow cytometer,    transporting the sample and the reagent to said thermal cycler for amplification, and    analyzing the sample with said flow cytometer utilize beads with each bead having a unique spectral address.    
     
     
         46 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing polystyrene latex microspheres beads with each bead having a unique spectral address.  
     
     
         47 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing beads with varying ratios of red and orange infrared dyes giving each bead a unique spectral address.  
     
     
         48 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing a 100-plex array of beads with varying ratios of red and orange infrared dyes giving each bead a unique spectral address.  
     
     
         49 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing beads with each bead having a capture antibody specific for a target antigen.  
     
     
         50 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing beads with at least one bead having an anthrax capture antibody specific for a target anthrax antigen.  
     
     
         51 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing beads with at least one bead having a plague capture antibody specific for a target plague antigen.  
     
     
         52 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing beads with at least one bead having a small pox capture antibody specific for a target small pox antigen.  
     
     
         53 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing beads with at least one bead having a botox capture antibody specific for a target botox antigen.  
     
     
         54 . The nucleic acid assay method of  claim 45  wherein said step of analyzing the sample with said flow cytometer comprises utilizing beads with each bead having a capture antibody specific for a target antigen and a fluorescent reporter.