Rapid microbial detection

Abstract

Devices and methods are provided to detect the presence of bacteria and small microorganisms, and to identify various microbial attributes rapidly.

1 . A method of processing within a microfluidic device a sample that includes microorganisms and non-microbial cells, the method comprising:
 selectively lysing material including non-microbial cells to form lysed debris within the sample while leaving intact microorganisms within the sample.   
     
     
         2 . The method of  claim 1 , further comprising:
 separating and removing lysed debris from non-lysed and intact microorganisms within the sample.   
     
     
         3 . The method of  claim 1 , further comprising:
 identifying the intact microorganisms within the sample.   
     
     
         4 . The method of  claim 2 , further comprising:
 identifying the intact microorganisms within the sample.   
     
     
         5 . The method of  claim 2 , further comprising collecting lysed debris separated from the sample in a waste collection stage of the microfluidic device. 
     
     
         6 . The method of  claim 2 , wherein the separating and removing lysed debris from non-lysed and intact microorganisms within the sample occurs without using a nucleotide, an antibody, or other ligand or reagent that specifically recognizes and binds to a microorganism. 
     
     
         7 . The method of  claim 1 , wherein the microorganisms are selected from the group consisting of bacteria, fungi, and small unicellular and multicellular organisms. 
     
     
         8 . The method of  claim 1 , wherein the microorganisms are selected from the group of genera consisting of  Staphylococcus, Escherichia, Listeria, Salmonella, Streptococcus, Klebsiella  and  Campylobacter.    
     
     
         9 . The method of  claim 1 , wherein the sample is a liquid selected from the group consisting of whole blood, plasma, serum, saliva, urine, cerebrospinal fluid, water, and fruit and vegetable juices. 
     
     
         10 . The method of  claim 1 , wherein the sample is selected from the group consisting of meat, produce, processed food, dairy products, poultry products, pharmaceutical process streams, bulk drug substance, and final drug product. 
     
     
         11 . The method of  claim 1 , wherein the sample has a volume ranging from about 0.1 mL to about 10 mL. 
     
     
         12 . The method of  claim 3 , wherein the identifying intact microorganisms within the sample comprises:
 subjecting the sample to infrared spectrometric analysis utilizing a detector; and   determining a presence of intact microorganisms within the sample based upon the infrared spectrometric analysis.   
     
     
         13 . The method of  claim 12 , wherein the identifying intact microorganisms within the sample further comprises:
 determining a species of one or more intact microorganisms within the sample based upon the infrared spectrometric analysis.   
     
     
         14 . The method of  claim 12 , wherein the determining a species of one or more intact microorganisms within the sample further comprises:
 providing data associated with the infrared spectrometric analysis to a computer device for determination of the species of the one or more intact microorganisms.   
     
     
         15 . The method of  claim 12 , wherein the determining a species of one or more intact microorganisms within the sample further comprises:
 comparing spectral data obtained from the infrared spectrometric analysis with spectral data from a database; and   identifying the species of the microorganism based upon the comparing.   
     
     
         16 . The method of  claim 3 , further comprising:
 prior to identifying the intact microorganisms within the sample, concentrating the microorganisms in the sample via reducing a volume of the sample and/or filtration of the sample.   
     
     
         17 . The method of  claim 1 , wherein the selective lysing is achieved by passing the sample through stationary microporous surfaces. 
     
     
         18 . The method of  claim 17 , wherein the microporous surfaces include pores having diameters from 1 micrometer to 10 micrometers. 
     
     
         19 . The method of  claim 1 , wherein the selective lysing is achieved by passing the sample through a porous monolithic structure including microporous surfaces. 
     
     
         20 . The method of  claim 1 , further comprising, prior to the selective lysing, separating and removing from the sample components that are larger in size than microorganisms within the sample. 
     
     
         21 . The method of  claim 20 , wherein the separation and removal from the sample components that are larger in size than microorganisms within the sample comprises passing the sample through a channel having a spiral path within the microfluidic device. 
     
     
         22 . The method of  claim 20 , wherein the separation of lysed debris from non-lysed and intact microorganisms within the sample comprises passing the sample through a linear array of posts within the microfluidic device to separate, via deterministic lateral displacement (DLD), lysed debris from the sample while maintaining the microbial cells of interest within the sample.