Separation device for use in the separation, characterization and/or identification of microorganisms
The present invention is directed to a separation device or container that can be used in the separation, isolation or pelleting of microorganisms from a test samples known to contain or suspected of containing said microorganisms. Subsequently, the separated, isolated or pelleted microorganism sample can undergo one or more interrogation steps to provide measurements useful for the characterization and/or identification of microorganism. In one aspect of the present invention, the interrogation steps can occur in situ in the separation device or container described herein.
1. A method comprising the steps of:
separating at least a portion of a liquid sample in a container, the container comprising:
a longitudinal axis;
an upper portion having an opening;
a lower portion extending from the opening, the lower portion having a generally cylindrical outer shape, the lower portion comprising
an internal chamber, the internal chamber comprising an upper reservoir, and a middle tapered section connecting the upper reservoir to a capillary tube of diameter smaller than that of the upper reservoir, the upper reservoir, the middle tapered section and the lower capillary tube arranged around the longitudinal axis of the container;
an optical window transparent to light and configured for interrogation of a portion of the capillary tube; and
a closure cap operably positioned about the opening;
pelletizing at least a portion of the liquid sample from the separating step so as to form an isolated, pelletized microorganism or contaminant sample;
interrogating at least a portion of the isolated, pelletized microorganism or contaminant sample from the pelletizing step; and
characterizing and/or identifying one or more of microorganisms or contaminants present in the isolated, pelletized microorganism or contaminant sample
wherein the separating step does not result in banding.
2. The method of claim 1 , wherein the liquid sample is a blood culture.
3. The method of claim 1 , wherein the container comprises a density cushion and the separating step comprises centrifuging the container for a sufficient time and/or force that substantially prevents a density gradient from forming within the density cushion.
4. The method of claim 1 , wherein the separating step does not result in isopycnic sedimentation.
5. The method of claim 1 , wherein the separating step and/or the pelletizing step is not complete prior to carrying out the interrogating step.
6. The method of claim 1 , wherein the container further comprises a lysing solution and the separating step comprises lysing the sample.
7. The method of claim 1 , wherein the interrogation step is carried out while the separated, isolated or pelleted microorganism sample is present in the container.
8. The method of claim 1 , wherein the interrogating step comprises a non-invasive interrogation technique.
9. The method of claim 1 , wherein the interrogating step is intrinsic fluorescence spectroscopy, Raman spectroscopy, or Mass Spectrometry.
10. A method comprising the steps of:
separating, without banding and isopycnic sedimentation, at least a portion of a liquid sample in a container, the container comprising:
a longitudinal axis;
an upper portion having an opening;
a lower portion extending from the opening, the lower portion having a generally cylindrical outer shape, the lower portion comprising
an internal chamber, the internal chamber comprising an upper reservoir, and a middle tapered section connecting the upper reservoir to a capillary tube of diameter smaller than that of the upper reservoir, the upper reservoir, the middle tapered section and the lower capillary tube arranged around the longitudinal axis of the container;
an optical window transparent to light and configured for the interrogation of a portion of the capillary tube;
a closure cap operably positioned about the opening; wherein the container comprises a density cushion; and
centrifuging the container for a sufficient time and/or force that substantially prevents a density gradient from forming within the density cushion;
pelletizing at least a portion of the liquid sample from the centrifuging step so as to form an isolated, pelletized microorganism or contaminant sample;
interrogating at least a portion of the isolated, pelletized microorganism or contaminant sample from the pelletizing step; and
characterizing and/or identifying one or more of microorganisms or contaminants present in the isolated, pelletized microorganism or contaminant sample.