Methods and apparatus for assays of bacterial spores

Abstract

Described herein are methods and apparatus for assays of bacterial spores. In particular, methods and apparatus for lateral flow immunoassay for bacterial spore detection and quantification, live/dead assay for bacterial spores, lifetime-gated measurements of bacterial spores and imaging bacterial spores using an active pixel sensor, and unattended monitoring of bacterial spores in the air are described.

1 - 27 . (canceled) 
     
     
         28 . A method for detection of bacterial spores in air comprising:
 collecting an air sample;   suspending the collected sample in a solution including lanthanide ions in the apparatus according to  claim 38 ;   treating the suspended sample to release DPA from the bacterial spores;   exciting combined lanthanide ions and DPA to generate a luminescence characteristic of the combined lanthanide ions and DPA; and   detecting luminescence to determine the presence of the bacterial spores.   
     
     
         29 . The method of  claim 28  wherein collecting an air sample comprises capturing the air sample with an aerosol sampler or impactor. 
     
     
         30 . The method of  claim 28  wherein detecting luminescence to determine the presence of the bacterial spores comprises monitoring the luminescence. 
     
     
         31 . The method of  claim 28  wherein the collecting comprises continuously sampling the air. 
     
     
         32 . The method of  claim 28 , wherein detecting the luminescence to determine the presence of the bacterial spores comprises continuously monitoring the luminescence. 
     
     
         33 . The method of  claim 28  wherein suspending the collected sample in a solution comprises cooling the heated solution to increase the fraction of bound Tb-DPA complex. 
     
     
         34 . The method of  claim 28  wherein treating the collected sample to release DPA from the bacterial spores comprises microwaving the bacterial spores to heat the solution. 
     
     
         35 . The method of  claim 28 , wherein treating the collected sample comprises cooling the heated solution to increase the fraction of combined lanthanide ion-DPA complex. 
     
     
         36 . The method of  claim 28 , further comprising generating an alarm signal when presence of bacterial spores is detected. 
     
     
         37 . The method of  claim 28 , further comprising generating an alarm signal when a concentration of bacterial spores reaches a predetermined magnitude. 
     
     
         38 . An apparatus for unattended monitoring of bacterial spores in air-comprising:
 a biosampler for collecting an air sample, the biosampler having a collection vessel containing a solution including lanthanide ions, the collection vessel configured to further include bacterial spores from a collected air sample suspended in the solution including lanthanide ions;   means for releasing DPA from the suspended bacterial spores in the solution to allow the DPA to combine with the lanthanide ions to form a lanthanide-DPA complex in the collection vessel containing the solution comprising the suspended bacterial spores;   an energy source for exciting the lanthanide-DPA complex to generate luminescence; and   an electro-optical circuit comprising a luminescence detector configured to measure the luminescence generated by the lanthanide-DPA complex.   
     
     
         39 . The apparatus of  claim 38 , further comprising an alarm circuit coupled to the electro-optical circuit to detect a bacterial spore concentration above a predetermined threshold. 
     
     
         40 . A method for spore detection, the method comprising
 combining collected samples with lanthanide ions in a solution in the apparatus according to  claim 48 ;   treating the solution comprising the collected sample to release DPA from bacterial spores   detecting luminescence from the solution to detect bacterial spores.   
     
     
         41 . The method of  claim 40 , wherein the combining is performed by suspending the collected sample in a solution, providing terbium ions in the solution and treating the solution to release DPA bacterial spores. 
     
     
         42 . The method of  claim 40 , wherein the treatment comprises microwaving or sonicating the solution. 
     
     
         43 . The method of  claim 40 , wherein treating the collected sample comprises cooling the heated solution to increase the fraction of combined lanthanide-DPA complex. 
     
     
         44 . The method of  claim 40 , wherein the detecting is performed by detecting luminescence intensity to quantify presence of bacterial spores in the solution. 
     
     
         45 . The method of  claim 40 , wherein the detecting comprises continuously monitoring the luminescence. 
     
     
         46 . The method of  claim 40 , further comprising generating an alarm signal when presence of bacterial spores is detected. 
     
     
         47 . The method of  claim 40 , further comprising generating an alarm signal when a concentration of bacterial spores reaches a predetermined magnitude. 
     
     
         48 . An apparatus for monitoring of bacterial spores the apparatus comprising:
 a collection vessel containing a solution including lanthanide ions, thef collection vessel configured to further include bacterial spores from a collected sample suspended in the solution including lanthanide ions;   means for releasing DPA from the suspended bacterial spores in the solution to allow the DPA to combine with the lanthanide ions to form a lanthanide-DPA complex in the collection vessel containing the solution comrising the suspended bacterial spores;   an energy source for exciting the lanthanide-DPA complex to generate luminescence; and   a circuit comprising a luminescence detector configured to measure the luminescence generated by the lanthanide-DPA complex.   
     
     
         49 . The apparatus of  claim 48 , further comprising an alarm circuit coupled to the circuit to detect a bacterial spore concentration above a predetermined threshold. 
     
     
         50 . The apparatus of  claim 48 , wherein the means for releasing DPA comprises a microwave.