Patent ID: 8647835
Filing Date: 2014-02-11
Classification: C12Q,G01N

Abstract:
1. A method of identifying two or more different and unknown microorganism species from a test sample, comprising: (a) obtaining a test sample known to contain or that may contain two or more different, unknown microorganism species; (b) optionally selectively lysing non-microorganism cells in said test sample to produce a lysed sample; (c) layering said test or lysed sample on a density cushion in a container wherein said density cushion has a homogenous density of from about 1.025 to about 1.22 g/ml; (d) centrifuging the container to separate said two or more microorganism species from other components of said sample, said microorganism species passing through said density cushion to form a mixed pellet of said two or more microorganism species at the bottom of said container, wherein said two or more microorganism species form two or more enriched layers within said mixed pellet; (e) spectroscopically interrogating each of said enriched layers of said two or more microorganism species in said mixed pellet in situ to produce excitation-emission matrix (EEM) of said two or more unknown microorganism species, wherein said spectroscopic interrogation comprises intrinsic fluorescence, and wherein said intrinsic fluorescence is measured in front face mode; and (f) identifying each of said two or more unknown microorganism species in said enriched layers of said mixed pellet by comparison of the spectroscopic measurements with spectroscopic measurements taken, or spectroscopic properties predicted, of known microorganisms, wherein said two or more unknown microorganisms are identified to the genus level, species level, and/or strain level.