System for Conducting the Identification of Bacteria in Urine
20200240977 ยท 2020-07-30
Inventors
- Gal Ingber (Oranit, IL)
- William G. Atterbury (Columbus, OH)
- Russell H. Barnes (Columbus, OH, US)
- Douglas E. Boyd (Columbus, OH, US)
- Joseph D. Dennis (Granview Heights, OH, US)
- Jonathan Gurfinkel (Omer, IL)
- Dave Holley (Lancaster, OH, US)
- Steven E. Huckaby (Blacklick, OH, US)
- Thomas A. KLAUSING (Powell, OH, US)
- John S. Laudo (Columbus, OH, US)
- Kevin Sadeski (Canal Winchester, OH, US)
- Jason A. Schaefer (Coral Springs, FL, US)
- K. Bryan Scott (Westerville, OH, US)
- Carol Stillman (Powell, OH, US)
- Sherwood Talbert (Columbus, OH, US)
- John Tallarico (Powell, OH, US)
Cpc classification
G01N35/025
PHYSICS
G01N21/0303
PHYSICS
C12Q1/04
CHEMISTRY; METALLURGY
International classification
C12Q1/04
CHEMISTRY; METALLURGY
Abstract
A system for conducting the identification and quantification of micro-organisms, e.g., bacteria in urine samples which includes: 1) several disposable cartridges for holding four disposable components including a centrifuge tube, a pipette tip having a 1 ml volume, a second pipette tip having a 0.5 ml volume, and an optical cup or cuvette; 2) a sample processor for receiving the disposable cartridges and processing the urine samples including transferring the processed urine sample to the optical cups; and 3) an optical analyzer for receiving the disposable cartridges and configured to analyze the type and quantity of micro-organisms in the urine sample. The disposable cartridges with their components including the optical cups or cuvettes are used in the sample processor, and the optical cups or cuvettes containing the processed urine samples are used in the optical analyzer for identifying and quantifying the type of micro-organism existing in the processed urine samples.
Claims
1-16. (canceled)
17. A method for processing a fluid sample in preparation for the identification and quantification of micro-organisms in the fluid sample, the steps comprising: a) obtaining a fluid sample; b) passing the fluid sample through a filter; c) obtaining a sample of the filtered fluid sample and placing it into a centrifuge tube; d) obtaining a dilution of the dissolved material in the fluid sample while retaining bacteria in the fluid sample by performing the following steps: 1) centrifuging the sample of step c); 2) decanting a portion of the fluid in the centrifuge tube of step 1); 3) replacing the decanted solution of step 2) with a saline solution; and e) transferring the final solution of step 4) into an optical cup or cuvette for analysis.
18. The method of claim 17, wherein the fluid sample is urine.
19. The method of claim 17, wherein step d) results in about a 1,000,000:1 dilution.
20. The method of claim 17, wherein the sample of step (d)(1) is centrifuged at about 12,000 g-forces.
21. The method of claim 17, wherein about 95% of the fluid from step (d)(2) is decanted.
22. The method of claim 17, wherein after step d(3), further comprising a step of repeating steps (1), (2) and (3) and then transferring the final solution into an optical cup or cuvette for analysis.
23. The method of claim 17, wherein after step (d)(3), further comprising a step of repeating steps (1), (2) and (3) for a plurality of times and then transferring the final solution into an optical cup or cuvette for analysis.
24. The method of claim 23, wherein the plurality of times is at least five times.
25-37. (canceled)
38. A method for identifying the type and quantity of micro-organisms in a fluid sample, the steps comprising: a) obtaining a fluid sample; b) passing the fluid sample through a filter; c) obtaining a sample of the filtered fluid and placing it into a centrifuge tube; d) obtaining a dilution of the dissolved material in the urine sample while retaining bacteria in the urine sample by performing the following steps: 1) centrifuging the sample of step c); 2) decanting a portion of the fluid in the centrifuge tube of step 1); 3) replacing the decanted solution of step 2) with a saline solution; and e) transferring the final solution of step 4) into an optical cups or cuvette; and f) subjecting the optical cup or cuvette to an optical analysis having optics which include exciting the fluid sample with a plurality of different wavelengths, collecting and detecting the fluorescent emissions; and directing the fluorescent emissions into a spectrometer.
39. The method of claim 38, wherein step d) results in about a 1,000,000:1 dilution.
40. The method of claim 38, wherein the sample of step (d)(1) is centrifuged at about 12,000 g-forces.
41. The method of claim 38, wherein about 95% of the fluid from step (d)(2) is decanted.
42. The method of claim 38, wherein after step d(3), further comprising a step of repeating steps (1), (2) and (3) and then transferring the final solution into an optical cup or cuvette for analysis.
43. The method of claim 38 wherein after step (d)(3), further comprising a step of repeating steps (1), (2) and (3) for a plurality of times and then transferring the final solution into an optical cup or cuvette for analysis.
44. The method of claim 43 wherein the plurality of times is at least five times.
45. A method for identifying the type and quantity of micro-organisms in a fluid sample, the steps comprising: a) obtaining a fluid sample; b) passing the fluid sample through a filter; c) obtaining a sample of the filtered fluid and placing it into a removable tube; and d) transferring the sample of the filtered fluid into an optical cup or cuvette, said optical cup or cuvette comprising a container having an open portion and a sidewall that is continuously tapered along their length to a truncated conical end and a reflective surface; and e) subjecting the optical cup or cuvette to an optical analysis having optics which include exciting the fluid sample with a plurality of different wavelengths, collecting and detecting the fluorescent emissions, and directing the fluorescent emissions into a spectrometer, wherein during the optical analysis, a light is directed down a middle portion of the container through the open portion of the container and into the fluid sample, wherein the sidewall is continuously tapered along its length to a truncated conical end configured to assist with the optical analysis and the reflective surface of the container reflects fluorescent emissions back in an upwardly direction out of the fluid sample and through the open portion of the container to an optics system for the optical analysis.
46. The method of claim 45, wherein the container includes a lip for suspending the container from a top surface of a cartridge.
47. The method of claim 46, wherein the cartridge includes a body and the container is located within an opening in the cartridge body.
48. The method of 45, wherein the container has a flat bottom surface.
49. The method of claim 45, wherein the reflective surface is a coated onto the container or is a separately formed layer of material.
50. The optical cup or cuvette of claim 45, wherein the reflective surface is an inner reflective surface of the container or is incorporated onto the container material.
Description
BRIEF DESCRIPTION OF THE DRAWINGS
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DETAILED DESCRIPTION OF THE INVENTION
[0024] The present invention will be described with reference to the accompanying drawings where like reference numbers correspond to like elements.
[0025] Referring to
[0026] The centrifuge tube 18 is a container that has an elongated body 18b with a tapered end indicated at 18a. In general, the centrifuge tube 18 initially contains the urine sample and the first pipette tip 20 may be used to dilute the urine-dissolved constitutes, and the second pipette tip 24 may be used to transfer the diluted urine sample into the optical cup or cuvette 22 for optical analysis. The disposable cartridge 12 and its disposable components 18, 20, 22, and 24 may be made of a plastic material which is easily molded and inexpensive to manufacture.
[0027] Still referring to
[0028] Referring to
[0029] Several disposable cartridges 12 each containing the four disposable components 18, 20, 22, and 24 are then inserted into a magazine 26 shown at the top of
[0030] The sample processor 14 of
[0031] In general, centrifuge tube 18 contains about 2 ml sample of filtered urine which is placed into the centrifuge tube by the user. This sample may then be sufficiently diluted with a saline solution or water by centrifuging the sample followed by using the first pipette tip 20 with the 1.0 ml volume to decant the supernates in two decant cycles followed by refilling of the centrifuge tube 18 with a saline or water. The second pipette tip 24 having the 0.5 ml volume may then be used to draw out about 500 l of fluid from centrifuge tube 18 and then to dispense this 500 l of fluid into the respective optical cup or cuvette 22 of the designated patient. This second pipette tip 24 can then be inserted into the first pipette tip 20 and both pipette tips 20, 24 can be disposed of properly. It is believed that one pipette tip may be used to dilute and draw out instead of two pipette tips. This process may be done manually or may be done automatically.
[0032] The loading and unloading of the magazine 26 is accomplished with the several disposable cartridges 12 mounted on the rotatable table 40 (
[0033] The transfer of the centrifuge tube 18 (
[0034] Centrifuge 30 (
[0035] There are two fluid transfer arms 34, 34a (
[0036] The syringe pump dispenser fluid system 36, is illustrated in
[0037] After the sample in centrifuge tube 18 has been sufficiently diluted with the clean fluid, one of the two fluid transfer arms 34, 34a (
[0038] The metering/decanting, metering/refilling, and metering/fluid transferring process described herein above is to obtain preferably, approximately a 1,000,000:1 dilution of the dissolved materials retaining bacteria in the urine sample in centrifuge tube 18. This can be achieved by 1) centrifuging, through means known to those skilled in the art, the urine sample at a 12,000 g-force; 2) decanting about 95% of the fluid by using the first pipette tip 20; 3) replacing the decanted solution of 2) with a saline solution; and 4) repeating steps 1), 2), and 3) at least five times by using the first pipette 20. The final processed urine sample in centrifuge tube 18 can then be decanted via the second pipette 24 into the optical cup or cuvette 22.
[0039] The final processed urine sample in optical cup or cuvette 22 can then be used in an optical analysis for determining the micro-organism's identity and/or quantity in the urine sample in optical cup or cuvette 22. This information can be obtained by using the system as disclosed in the aforesaid U.S. Publication No. 2007/0037135 A1.
[0040] Each of the steps described above for one centrifuge tube 18 is done in the sample processor 14 for each of the disposable cartridges 12 in magazine 26. It is to be appreciated that the waste fluid of each disposable cartridge 12 is disposed into a receptacle (not shown) in sample processor 14 or is plumbed directly into a drain. The waste disposables, i.e., the disposable cartridge 12 and disposable components 18, 20, 22, and 24 remain on the magazine 26 for manual removal when the magazine 26 is unloaded in preparation for the next operation of the sample processor 14 for processing the next batch of urine samples.
[0041] The following steps are involved in processing the urine samples in preparation for analysis via the optical analyzer 16 of
[0042] The optical analyzer 16 of
[0043] As can be appreciated, a cartridge 12 that has the cups or cuvettes 22 containing the processed urine sample for optical analysis are placed into the holders 56 of the magazine 54.
[0044]
[0045]
[0046] The optics system 44 of the optical analyzer 16 will now be described. The optics system is shown in greater detail in
[0047] In addition, the optical collection unit includes optical elements to gather and direct the fluorescent emissions of the samples in the cups or cuvettes 22 into the spectrometer.
[0048] The optics system 44 may include a Czerny-Turner spectrometer with a CCD (charged couple device) Photon Detector, whereby fluorescent photons are reflected by several mirrors before contacting the CCD device. The emitted fluorescence will be monitored on the CCD device by integrating for a period of time. It is also envisioned that the Czerny-Turner spectrometer be modified with additional cylindrical lenses adjacent the entrance slit and the CCD device in order to improve photon usage efficiency. Additionally, as schematically illustrated in
[0049] Referring to
[0050] Both the sample processor 14 and the optical analyzer 16 will have a HEPA air-filtering system for ventilation purposes in filtering the air entering and exiting the sample processor 14 and the optical analyzer 16. The sample processor 14 and the optical analyzer 16 will also have a slight positive pressure during operation. However, this pressure will decrease to atmospheric pressure when the user is allowed to open the door. Power to both the sample processor 14 and the optical analyzer 16 will be supplied from a separate rack mounted power supply module.
[0051] It is further envisioned that the LED intensity will be monitored to correlate the emitted fluorescence with the intensity of the excitation fluorescence. In particular, the information obtained by the optical analyzer 16 may be used to generate graphs similar to FIGS. 5 through 9 of U.S. Publication No. 2007/0037135 A1, described in greater detail below. The graphs represent for the concentration of the bacteria in the sample cups or cuvettes 22, the fluorescence intensity, the emission wavelengths and the excitation wavelengths.
[0052] As discussed hereinabove, the optical analyzer 16 provides results that are then used to identify the type of bacteria in the urine samples. This can be done by coupling the optical analyzer 16 to a computer module (not shown) and feeding in the acquired information of the optical analyzer 16, such as the fluorescence emission, into the computer module. The computer module may perform multivariate analysis on the fluorescence excitation-emission matrices of the urine samples to identify and quantify the urine samples in a manner similar to that disclosed in the above U.S. Publication No. US 2007/0037135 A1. Here, the system includes a fluorescence excitation module which includes an excitation light source, a sample interface module for positioning the sample to receive the light source, a fluorescence emission module and a detection device. The computer module described above is coupled to the fluorescence module. The multivariate analysis may comprise extended partial least squared analysis for identification and quantification of the urine samples.
[0053] It is still further envisioned that a homogenitor tube will be used to mix the different LED packages output into a uniform UV light source. A typical homogenitor tube for use in the invention will be similar to that known to those skilled in the art.
[0054] The present invention has been described with reference to the preferred embodiments. Obvious modifications and alterations will occur to others upon reading and understanding the preceding detailed description. It is intended that the invention be construed as including all such modifications and alterations.