DNA ARRAY ANALYSIS AS A DIAGNOSTIC FOR CURRENT AND EMERGING STRAINS OF INFLUENZA

• US 20090124512A1
• Filed: 01/18/2007
• Published: 05/14/2009
• Est. Priority Date: 01/18/2006
• Status: Abandoned Application

• Current Assignee: Centers For Disease Control and Prevention (Government of the United States of America), The Board of Regents of the University of Colorado (University of Colorado System)
• Original Assignee: The Board of Regents of the University of Colorado (University of Colorado System)
• Inventors: Mehlmann, Daniela, Smith, Catherine B., Moore, Chad L., Cox, Nancy, Rowlen, Kathy L., Townsend, Michael, Kuchta, Robert, Mehlmann, Martin, Smagala, James, Dawson, Erica
• Application Number:
  US12/160,784
• Publication Number:
  US 20090124512A1
• Time in Patent Office: Days
• US Class Current: 506/9
• CPC Class Codes: C12Q 1/701 Specific hybridization probes
• CPC Class Codes: C12Q 2565/501 being an array of oligonucleotides

Abstract

Embodiments herein provide for methods, compositions and apparati for detection and/or diagnosis of virus types, subtypes and/or strains. In particular embodiments, the virus is an influenza virus. The apparatus may include a microarray with attached capture probes, designed to bind to oligonucleotides capable of binding at least a portion of a nucleic acid sequence of one or more target genes in a broad array of influenza types, subtypes or strains. The compositions may include isolated nucleic acids as capture probes, target sequences and/or tagged label probes, of use for diagnosis and/or detection of influenza virus.

Claims

1. An array comprising:

a plurality of capture probes comprising oligonucleotides, wherein the capture probes are capable of binding to oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene of one or more influenza virus.

2. The array of claim 1, wherein the capture probes are capable of binding to one or more influenza type.

3. The array of claim 1, wherein the capture probes are capable of binding to one or more influenza A subtype or strain.

4. The array of claim 1, wherein the plurality of capture probes are bound to the surface of a solid substrate.

5. The array of claim 4, wherein the array contains 100 or less capture probes bound to the surface of the solid substrate.

6. The array of claim 4, wherein the solid substrate is selected from the group consisting of glass, plastic, silicon-coated substrate, macromolecule-coated substrate, particles, beads, microparticles, microbeads, dipstick, magnetic beads, paramagnetic beads and a combination thereof.

7. The array of claim 4, further comprising a positive control probe bound to the surface of the solid substrate, wherein the positive control probe is capable of indicating conditions sufficient to form a complex of a capture probe binding to an oligonucleotide comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene.

8. The array of claim 1, wherein the array is a microarray.

9. The array of claim 8, wherein the microarray is a multiplex characteristic array derived from more than one target gene.

10. The array of claim 1, wherein the capture probes are capable of binding to an influenza strain selected from the group consisting of influenza A H3N2, influenza A H1N1, influenza A H5N1, influenza A H7N7, influenza A H9N2, influenza A H3N8, influenza A H1N2, influenza A H3N3, influenza A H3 and a combination thereof.

11. The array of claim 1, wherein the oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence are derived from a single target gene segment.

12. The array of claim 1, wherein the capture probes are selected from sequences listed in Table 3, Table 4, or a combination thereof.

13. The array of claim 1, wherein each of the capture probes are independently about 10 to about 100 nucleotides (nt) in length.

14. A method for producing an array for detecting the presence of influenza virus comprising:

attaching a plurality of capture probes to a solid substrate surface to form an array, wherein the capture probes are capable of binding to oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene of one or more influenza virus.

15. The method of claim 14, wherein the capture probes are capable of binding to one or more influenza type.

16. The method of claim 14, wherein the capture probes are capable of binding to one or more influenza A subtype or strain.

17. The method of claim 14, further comprising binding a positive control probe to the surface of the solid substrate, wherein the positive control probe is capable of indicating conditions sufficient to form a complex of a capture probe binding to an oligonucleotide comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene.

18. The method of claim 14, wherein the capture probes are capable of binding to oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene selected from the group consisting of hemagglutinin (HA gene segment), neuraminidase (NA gene segment), matrix protein (M gene segment) and a combination thereof.

19. The method of claim 14, wherein the capture probes are capable of binding to oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of the M gene segment.

20. A method for detecting influenza in a sample, the method comprising:

a) contacting the sample with an array of a plurality of capture probes to produce a test array, wherein the test array comprises a capture probe-sample complex when the sample contains an oligonucleotide comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene of one or more influenza virus; and

b) contacting the test array with one or more detection probes to produce a labeled array, wherein the labeled array comprises a target-probe complex when the test array comprises the a capture probe-sample complex, and wherein the presence of the target-probe complex is indicative of the presence of influenza virus in the sample.

21. The method of claim 20, wherein the array comprises a plurality of capture probes comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of one or more target genes of at least one influenza virus type, subtype or strain.

22. The method of claim 20, wherein the presence of influenza virus in the sample is determined by detecting a signal generated by the probe of a target-probe complex.

23. The method of claim 22, wherein the signal generated by the target-probe complex produces different patterns depending on the influenza type, subtype or strain present in the sample.

24. The method of claim 20, wherein the capture probes are capable of binding to one or more influenza type.

25. The method of claim 20, wherein the capture probes are capable of binding to one or more influenza A subtype or strain.

26. The method of claim 20, further comprising a positive control probe bound to the surface of the solid substrate, wherein the positive control probe is capable of indicating conditions sufficient to form a complex of a capture probe binding to an oligonucleotide comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene.

27. The method of claim 20, further comprising a negative control probe bound to the surface of the solid substrate, wherein the negative control probe is capable of indicating conditions sufficient to indicate specificity of the capture label probes to bind to influenza virus and not to the negative control probe.

28. The method of claim 20, wherein the target gene is selected from the group consisting of hemagglutinin (HA gene segment), neuraminidase (NA gene segment), matrix protein (M gene segment) and a combination thereof.

29. The method of claim 20, wherein the sample is selected from the group consisting of nasopharangeal washes, expectorate, optical swab, respiratory tract swabs, throat swabs, nasal swabs, nasal mucus, tracheal aspirates, bronchoalveolar lavage, mucus, blood, urine, tissue, saliva, air samples, air-filter samples, surface-associated samples and a combination thereof.

30. The method of claim 20, further comprising identifying the presence of influenza in a sample in 12 hours or less.

31. An array comprising:

a plurality of capture probes comprising oligonucleotides, wherein the capture probes are capable of binding to oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a single target gene segment of one or more influenza virus.

32. The array of claim 31, wherein the capture probes are capable of binding to one or more influenza type.

33. The array of claim 31, wherein the capture probes are capable of binding to one or more influenza A subtype or strain.

34. The method of claim 31, wherein the capture probes are capable of binding to oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of the M gene.

35. The method of claim 31, wherein the portion of a nucleic acid sequence or complimentary nucleic acid sequence of a single target gene segment comprise conserved sequence regions for a type, subtype or strain of influenza.

36. A kit comprising:

(a) an array of a plurality of capture probes bound to the surface of a solid substrate, wherein the capture probes are capable of binding to oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene of one or more influenza virus; and

(b) one or more tagged label probes wherein the tagged label probes are capable of producing a signal and wherein the label probes are capable of binding to the oligonucleotides comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene of one or more influenza virus.

37. The kit of claim 36, further comprising a positive control probe bound to the surface of the solid substrate, wherein the positive control probe is capable of indicating conditions sufficient to form a complex of a capture probe binding to an oligonucleotide comprising at least a portion of a nucleic acid sequence or complimentary nucleic acid sequence of a target gene.