Claims
- 1. A method for detecting the presence of a non-denatured nucleic acid analyte in a buffered test sample, said test sample consisting of suitable amounts of a blocking agent, and non-ionic detergent, said method comprising the steps of:
- (i) contacting the test sample with a first end of a test strip comprising a chromatographic material capable of moving the test sample laterally from the first end to a second end of the test strip by capillary migration and at least one single stranded capture nucleic acid molecule irreversibly affixed to the test strip at a specific capture zone, the capture nucleic acid being complementary to a portion of the nucleic acid analyte;
- (ii) incubating the test strip and test sample of step (i) for at least 5 minutes, whereby the test sample is allowed to traverse at least a portion of the test strip whereby the nucleic acid analyte is captured by hybridization to the capture nucleic acid in the capture zone;
- (iii) contacting the first end of the test strip with a suitable amount of wash buffer whereby unreacted nucleic acid analyte is removed from the capture zone;
- (iv) contacting the test strip with a signal-generating substance such that the signal-generating substance reacts with a reporter conjugate that is located either within the nucleic acid analyte or bound to the nucleic acid analyte to produce a detectable signal at the capture zone; and
- (v) comparing the detectable signal at the capture zone with a signal detectable at a portion of the test strip other than at the capture zone.
- 2. A method for detecting the presence of a non-denatured, randomly amplified polymorphic nucleic acid analyte in a test sample, comprising the steps of:
- (i) contacting the test sample with a first end of a test strip comprising a chromatographic material, the test strip having a specific capture zone in which the randomly amplified polymorphic nucleic acid analyte is immobilized;
- (ii) contacting the test strip with a hybridization solution consisting essentially of suitable amounts of a blocking agent, and non-ionic detergent and containing a detection probe, the detection probe comprising a base sequence complementary with the randomly amplified polymorphic nucleic acid analyte;
- (iii) incubating the test strip and test sample of step (ii) for at least five minutes whereby the test sample is allowed to traverse at least a portion of the test strip, whereby the detection probe and the randomly amplified polymorphic nucleic acid analyte hybridize;
- (iv) contacting the first end of the test strip with a suitable amount of wash buffer whereby the unreacted detection probe is removed from the capture zone;
- (v) contacting the test strip with a signal generating substance such that the signal generating substance reacts with a reporter conjugate located either within the detection probe or reversibly affixed to the test strip to produce a detectable signal at the capture zone; and
- (vi) comparing the detectable signal at the capture zone with a signal detectable at a portion of the test strip other than at the capture zone.
- 3. The method of claim 1 wherein said one single stranded capture nucleic acid molecule is irreversibly affixed to the test strip by means of a capture moiety.
- 4. The method of claim 1 where at step (iv) the reporter conjugate comprises a radioactive molecule linked to a member of a binding pair wherein said contacting is accomplished in the absence of a signal generating substance.
- 5. The method of claim 1 wherein the nucleic acid analyte comprises at least one reactive ligand.
- 6. The method of claim 1 wherein the reporter conjugate comprises a reporter moiety linked to a member of a binding pair, the reporter moiety selected from the group consisting of an enzyme, a chemiluminescent molecule, particles, and a fluorescent molecule.
- 7. The method of claim 6 wherein the reporter moiety is linked to an affinity-reactive member of a binding pair.
- 8. The method of claim 6 wherein the reporter moiety is linked to an immuno-reactive member of a binding pair, selected from the group consisting of antibodies antigens, haptens and anti-haptens.
- 9. The method of claim 6 wherein the reporter moiety is linked to a nucleic acid molecule capable of hybridizing to the nucleic acid analyte to be detected.
- 10. The method of claim 3 wherein the capture moiety is an affinity-reactive member of a binding pair.
- 11. The method of claim 3 wherein the capture moiety is an immuno-reactive member of a binding pair.
Parent Case Info
This application is a continuation-in-part of application Ser. No. 08/863,265, filed on May 27, 1997, abandoned, which is a file wrapper continuation of 08/530,795, filed on Sep. 20, 1995, abandoned, which is a file wrapper continuation of Serial No. 08/221,769, filed on Mar. 31, 1994, abandoned.
US Referenced Citations (7)
Foreign Referenced Citations (2)
Number |
Date |
Country |
2034313 |
Jul 1991 |
CAX |
0 170 746 |
Feb 1986 |
EPX |
Continuations (2)
|
Number |
Date |
Country |
Parent |
530795 |
Sep 1995 |
|
Parent |
221769 |
Mar 1994 |
|
Continuation in Parts (1)
|
Number |
Date |
Country |
Parent |
863265 |
May 1997 |
|