Self-contained device integrating nucleic acid extraction amplification and detection

• US 5,955,351 A
• Filed: 07/12/1996
• Issued: 09/21/1999
• Est. Priority Date: 07/13/1995
• Status: Expired due to Term

• Current Assignee: Applied Biosystems LLC (Thermo Fisher Scientific Incorporated)
• Original Assignee: Xtrana Inc. (Bio-Techne Corporation)
• Inventors: Kozwich, Diane L., Gerdes, John C., Jankovsky, Lynn D.
• Primary Examiner(s): Jones, W. Gary
• Assistant Examiner(s): WHISENANT, ETHAN C
• Application Number:
  US08/679,522
• Time in Patent Office: 1,166 Days
• Field of Search: 435/287.2, 435/6, 435/91.2, 422/68.1, 422/112, 436/810, 935/76, 935/77, 935/78
• US Class Current: 435/287.2
• CPC Class Codes: B01L 2300/047 Additional chamber, reservoir
• CPC Class Codes: B01L 2300/0663 Whole sensors
• CPC Class Codes: B01L 2300/0681 Filter
• 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 3/5082 Test tubes per se
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick

Abstract

A self-contained device is described that integrates nucleic acid extraction, specific target amplification and detection into a single device. This integration permits rapid and accurate nucleic acid sequence detection. The invention may be used, for example, in the screening for nucleic acid sequences which may be indicative of genetic defects or contagious diseases, as well as for monitoring efficacy in the treatment of contagious diseases.

Claims

1. A self-contained device for the extraction, amplification and detection of nucleic acid sequences, which comprises:

a) a first hollow elongated cylinder closed at one end having a plurality of chambers, each chamber having an upper end and a lower end;

b) a second hollow elongated cylinder positioned contiguously inside said first cylinder and having an upper end and a lower end with an aperture and sealing lip interposed and connecting via rotation said lower end of the second cylinder to said upper end of each chamber of said first cylinder, said second cylinder further having three indexing notches disposed equilaterally on the upper end of the cylinder; and

c) a cover integrally hinged to the open end of the first cylinder, said cover having a reaction bead chamber integral with a knife-edge, said chamber housing a reaction bead and hermetically sealed with a membrane, said cover further having an indexing pin disposed diametrically to the hinge for indexing with said notches during rotation of said first cylinder in relation to said second cylinder.

2. The device of claim 1 wherein the second hollow elongated cylinder further comprises extraction means and amplification means.

3. The device of claim 2 wherein the extraction means comprises dry lysing reagent for nucleic acid extraction.

4. The device of claim 2 wherein the amplification means comprises polymerases or ligases.

5. The device of claim 1 wherein said plurality of chambers of said first hollow elongated cylinder comprises a reservoir chamber and a detection chamber.

6. The device of claim 5 wherein said reservoir chamber is defined by the contiguous sides of the first hollow elongated cylinder, said detection chamber, and a porous membrane, said porous membrane having pores of a size to enable waste fluid to pass through.

7. The device of claim 5 wherein said detection chamber further comprises detection means.

8. The device of claim 7 wherein said detection means comprises an absorbent pad and a strip having colored microparticles and capture zones.

9. The device as defmed in claim 1, wherein the nucleic acid sequence to be amplified is any specific nucleic acid sequence.