Method for analyzing a fluid sample

• US 20010012612A1
• Filed: 03/06/2001
• Published: 08/09/2001
• Est. Priority Date: 05/28/1999
• Status: Active Grant

• Current Assignee: Cepheid (Danaher Corporation)
• Original Assignee: Cepheid (Danaher Corporation)
• Inventors: Chang, Ronald, Petersen, Kurt E., Northrup, M. Allen, Belgrader, Phillip, McMillan, William A., Pourahmadi, Farzad, Ching, Jesus, Dority, Douglas B., Sakai, Stanley H., Taylor, Michael T.
• Granted Patent: US 6,881,541 B2
• Time in Patent Office: Days
• US Class Current: 435/5
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0654 Lenses; Optical fibres
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0809 rectangular shaped
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0622 distribution valves, valves having multiple inlets and/or outlets, e.g. metering valves, multi-way valves
• CPC Class Codes: B01L 2400/0644 rotary valves
• CPC Class Codes: B01L 3/502 with fluid transport, e.g. in multi-compartment structures centrifugal-type cuvettes G01N21/07; analysis by separation into components G01N30/00; automatic analysers G01N35/00
• CPC Class Codes: B01L 7/52 with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples amplification or hybridisation processes per se C12Q1/68; controlling sequential reactions for synthesis B01J19/0046
• CPC Class Codes: C12N 1/066 by physical methods
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 436/805 Optical property
• CPC Class Codes: Y10S 436/806 Electrical property or magnetic property
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10S 436/829 Liposomes, e.g. encapsulation
• CPC Class Codes: Y10T 137/87716 For valve having a flexible diaphragm valving member
• CPC Class Codes: Y10T 137/88046 Biased valve with external operator
• CPC Class Codes: Y10T 436/255 : including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

A method for separating a desired analyte from a fluid sample comprises the steps of introducing the sample into a cartridge having a sample flow path and a lysing chamber in the sample flow path. The lysing chamber contains at least one filter for separating cells or viruses from the sample. The sample is forced to flow through the sample flow path, thereby capturing the cells or viruses with the filter as the sample flows through the chamber. The ratio of the volume of sample forced to flow through the chamber to the volume capacity of the chamber is preferably at least 2:1, and the volume of sample forced to flow through the chamber is preferably at least 100 microliters. The captured cells or viruses are disrupted to release the analyte therefrom, and the analyte is eluted from the chamber.

Claims

1. A method for separating a desired analyte from a fluid sample, the method comprising the steps of:

a) introducing the sample into a cartridge having;

i) a sample flow path; and

ii) a lysing chamber in the sample flow path, wherein the lysing chamber contains at least one filter for separating cells or viruses from the sample;

b) forcing the sample to flow through the sample flow path, thereby capturing the cells or viruses with the at least one filter as the sample flows through the chamber, wherein the ratio of the volume of sample forced to flow through the chamber to the volume capacity of the chamber is at least 2;

1, and wherein the volume of sample forced to flow through the chamber is at least 100 μ

l;

c) disrupting the captured cells or viruses to release the analyte therefrom; and

d) eluting the analyte from the chamber.

2. The method of

claim 1

, wherein the ratio of the volume of sample forced to flow through the chamber to the volume capacity of the chamber is at least 5:

1.

3. The method of

claim 1

, wherein the volume of sample forced to flow through the chamber is at least 1 ml.

4. The method of

claim 1

, wherein the step of disrupting the cells or viruses comprises agitating beads in the chamber.

5. The method of

claim 4

, wherein the beads have a binding affinity for the cells or viruses to be disrupted, and wherein the method further comprises the step of binding the cells or viruses to the beads.

6. The method of

claim 4

, wherein the beads have a binding affinity for the analyte, and wherein the method further comprises the step of binding the analyte to the beads.

7. The method of

claim 1

, wherein the lysing chamber contains a first set of beads for binding the cells or viruses and a second set of beads for rupturing the cells or viruses, the method further comprises the step of binding the cells or viruses to the first set of beads, and the step of disrupting the cells or viruses comprises rupturing the cells or viruses with the second set of beads.

8. The method of

claim 1

, wherein step of disrupting the cells or viruses comprises sonicating the chamber.

9. The method of

claim 1

, wherein step of disrupting the cells or viruses comprises heating the chamber.

10. The method of

claim 1

, further comprising the step of sonicating the chamber while forcing the sample to flow through the chamber.

11. The method of

claim 1

, wherein the step of eluting the analyte is preceded by the step of forcing a wash fluid to flow through the chamber while sonicating the chamber.

12. The method of

claim 1

, further comprising the step of sonicating the chamber while eluting the analyte.

13. The method of

claim 1

, wherein the cartridge has a first filter in the sample flow path for filtering coarse material from the sample and a second filter in the lysing chamber for separating the cells or viruses from the sample, the second filter having a smaller average pore size than the first filter, and wherein the step of forcing the sample to flow through the sample flow path comprises forcing the sample to flow through the first filter, thereby filtering the coarse material from the sample, prior to forcing the sample to flow through the second filter.