Apparatus and method for conducting an assay using reverse flow through a membrane

• US 5,788,863 A
• Filed: 12/13/1995
• Issued: 08/04/1998
• Est. Priority Date: 12/13/1995
• Status: Expired due to Fees

• Current Assignee: Becton, Dickinson & Co
• Original Assignee: Becton, Dickinson & Co
• Inventors: Milunic, David
• Primary Examiner(s): FORTUNA, ANA M
• Application Number:
  US08/571,572
• Time in Patent Office: 965 Days
• Field of Search: 210/651, 210/656, 210/198.2, 210/321.75, 210/321.84, 210/416.1, 210/321.72, 422/101
• US Class Current: 210/651
• CPC Class Codes: B01D 61/00 Processes of separation using semi-permeable membranes, e.g. dialysis, osmosis or ultrafiltration; Apparatus, accessories or auxiliary operations specially adapted therefor separation of gases or vapours by diffusion B01D53/22
• 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: G01N 33/54366 Apparatus specially adapted for solid-phase testing

Abstract

The present invention relates to an apparatus and method for conducting an assay using reverse flow through a membrane. More particularly, the present invention is directed to an apparatus and method which allows for the assaying of analytes, such as antigens, in a fluid sample containing such analytes with more accurate and reproducible results.

Claims

1. A device for separating an analyte from an analyte-containing fluid, said device comprising a body having a channel extending therethrough, said body including:

a fluid receiving portion;

a fluid movement influencing portion;

a membrane disposed between said fluid receiving portion and said fluid influencing portion capable of retaining an analyte from an analyte-containing fluid contacted therewith;

fluid moving means coupled with said fluid movement influencing portion capable of moving the fluid through said membrane in two directions;

wherein said membrane includes a capture phase; and

a detection component disposed in said device effective to interact with an analyte contained in the analyte-containing fluid forming an analyte detection component complex so as to render the analyte-detection component complex detectable.

2. The device according to claim 1, wherein said fluid moving means comprises a retractable plunger.

3. The device according to claim 1, wherein said analyte-containing fluid contains said detection component.

4. The device according to claim 3, wherein said detection component is group A streptococcus blue latex.

5. The device according to claim 1, wherein said membrane is constructed from nitrocellulose.

6. The device according to claim 1, wherein said analyte includes an antigen and said capture phase includes an antibody.

7. The device according to claim 6, wherein said antigen is group A streptococcus antigen and said antibody is group A streptococcus antibody.

8. The device according to claim 1 wherein the detection component is located in the fluid movement influencing portion.

9. The device according to claim 1 wherein the detection component is located in the channel.

10. The device according to claim 1 wherein the detection component is located in the fluid receiving portion.

11. The device according to claim 1 wherein the detection component is located in the fluid moving means.

12. A device for separating an analyte from an analyte-containing fluid, said device comprising:

a reservoir including at one end a fluid receiving portion capable of receiving a fluid and at the other end and analyte-retaining portion;

a hollow cylinder including at one end a portion adapted for joining said reservoir and said analyte-retaining portion and at the other end a portion dimensioned for receiving fluid moving means;

a membrane disposed between the analyte-retaining portion of said reservoir and the reservoir joining portion of said hollow cylinder;

fluid moving means dimensioned for retractable insertion within said hollow cylinder to cause movement of the fluid through said membrane in both directions; and

a detection component disposed within the device effective to interact with an analyte contained in the analyte-containing fluid forming an analyte-detection component complex so as to render the analyte detection component complex detectable.

13. A method for separating an analyte from an analyte-containing fluid, said method comprising the steps of:

(a) mixing within a reservoir an analyte containing fluid an a detection component effective to interact with an analyte contained in the analyte-containing fluid forming an analyte-detection component complex, so as to render the analyte-detection component complex detectable;

(b) contacting the analyte-detection component complex with a membrane disposed in said reservoir;

(c) passing the fluid in the reservoir though the membrane and into a chamber;

(d) passing the fluid back from the chamber through the membrane and into the reservoir, and thereafter through the membrane into the chamber;

(e) detecting any analyte-detection component complex retained on the membrane.

14. A reverse flow membrane assay kit comprising:

a device including a body having a fluid receiving portion and a fluid movement influencing portion;

a membrane disposed in said body having a first and second faces;

wherein said fluid movement influencing portion is capable of moving the fluid through said membrane in two directions with respect to said membrane faces;

a container in which is provided an amount of capture agent effective to form a capture phase with the membrane and interact with an analyte contained in an analyte-containing fluid so as to form an analyte-capture phase complex, and in which is provided an amount of detection component effective to interact with said analyte and said capture phase to form an analyte detection component complex, so as to render the analyte-detection component complex detectable.

15. A reverse flow membrane assay kit comprising:

a device including a body having a fluid receiving portion and a fluid movement influencing portion;

a membrane disposed within said fluid receiving portion;

wherein said fluid influencing portion is capable of moving the fluid in two directions with respect to the membrane;

a container in which is provided an amount of capture phase agent effective to form a capture phase with the membrane and interact with an analyte contained in an analyte-containing fluid so as to form an analyte-capture phase complex; and

a second container in which is provided an amount of detection component effective to interact with said analyte and said capture phase so as to form an analyte-detection component complex to render the analyte-detection component complex detectable.

16. A method of using a fluid withdrawing/dispensing device in communication with a diffuser containing a membrane to separate and analyte from an analyte-containing sample, said method comprising the steps of:

(a) introducing an analyte-containing fluid and a detection component in an amount of detection component effective to interact with an analyte contained in the analyte-containing fluid so as to form an analyte-detection component complex so as to render the analyte detection component complex detectable, into said fluid withdrawing/dispensing device;

(b) contacting the fluid with a membrane disposed in said device, and passing the fluid therethrough;

(c) passing the fluid back through the membrane in reverse direction; and

(d) detecting any analyte-detection component complex retained on the membrane.

17. The method according to claim 16, wherein said analyte includes an antigen and said capture phase includes an antibody.

18. The method according to claim 17, wherein said antigen is group A streptococcus antigen and said antibody is group A streptococcus antibody.

19. The method according to claim 18, wherein said detection component is group A streptococcus blue latex.