Methods and kits for the diagnosis of influenza
Some embodiments provided herein relate to combined assays. In some embodiments, an assay for identifying influenza type A or influenza type B is combined with an assay for determining the sensitivity of an influenza neuraminidase to an antiviral drug.
1. A diagnostic system for detecting influenza virus comprising:
a test for detecting influenza type A or type B in a sample, wherein the test is configured to process a sample comprising a first buffer compatible with the influenza type A or type B detection;
a cartridge for determining neuraminidase activity in the sample, wherein the cartridge is configured to process a sample comprising the first buffer into a sample comprising a second buffer, wherein the first buffer inhibits neuraminidase activity and is incompatible with the neuramidase determination, and wherein the second buffer is compatible with the neuraminidase determination; and
a first detector configured to measure a signal from the test and/or
the cartridge.
2. The system of claim 1 , wherein the cartridge comprises neuraminidase activity assay reagents selected from the group consisting of luciferase, a conjugate of a N-acetylneuraminic acid and luciferin, a derivative of a N-acetylneuraminic acid and luciferin conjugate, an antiviral drug, and a reagent comprising a masked inhibitor compound for use in a dual enzyme influenza neuraminidase sensitivity assay.
3. The system of claim 2 , wherein the masked inhibitor is selected from the group consisting of 1,1,1-Trifluro-4-(4-hydroxyphenyl)butan-2-one (para), 1,1,1-Trifluro-4-(3-hydroxyphenyl)butan-2-one (meta), and 1,1,1-Trifluro-4-(2-hydroxyphenyl)butan-2-one (ortho).
4. The system of claim 1 , wherein the cartridge comprises a matrix chamber comprising a cartridge matrix comprising the second buffer for the determination, and a reagent chamber comprising reagents for the neuraminidase activity determination.
5. The system of claim 4 , wherein the cartridge matrix comprises a cross-linked polysaccharide.
6. The system of claim 4 , wherein the matrix chamber and reagent chamber are in fluid communication such that a sample applied to the matrix chamber flows from the matrix chamber to the reagent chamber.
7. The system of claim 1 , wherein the cartridge comprises a multiwell cartridge adapted to be read using a photomultiplier tube or micro-photomultiplier tube.
8. The system of claim 1 , wherein the test comprises an immunoassay.
9. The system of claim 1 , wherein the immunoassay is a sandwich assay.
10. The system of claim 1 , wherein the test comprises a test matrix comprising the first buffer for the detection.
11. The system of claim 10 , wherein the test matrix comprises nitrocellulose.
12. The system of claim 1 , wherein the first buffer for the detection is lyophilized or dried down.
13. The system of claim 1 , wherein the first detector is configured to measure a signal from the test, and the system further comprises a second detector configured to measure a signal from the cartridge.
14. The system of claim 13 , wherein the first detector comprises a luminometer, and the second detector comprises a photomultiplier tube or micro-photomultiplier tube.
15. A kit for detecting influenza virus comprising:
a test for detecting influenza type A or type B in a sample, wherein the test is configured to process a sample comprising a first buffer compatible with the influenza type A or type B detection; and
a cartridge for determining neuraminidase activity in a sample, wherein the cartridge is configured to process a sample comprising an incompatible buffer for the determination to a sample comprising a compatible buffer for the determination, wherein the first buffer inhibits neuraminidase activity and is incompatible with the neuramidase determination, and wherein the second buffer is compatible with the neuraminidase determination.
16. The kit of claim 15 , further comprising reagents for a neuraminidase activity assay.
17. The kit of claim 16 , wherein the neuraminidase activity assay reagents are selected from the group consisting of luciferase, a conjugate of a N-acetylneuraminic acid and luciferin, a derivative of a N-acetylneuraminic acid and luciferin, a N-acetylneuraminic acid and a trifluoromethylketone, a derivative of a conjugate of a N-acetylneuraminic acid and a trifluoromethylketone, an antiviral drug, and a reagent comprising a masked inhibitor compound for use in a dual enzyme influenza neuraminidase sensitivity assay.
18. The kit of claim 17 , wherein the masked inhibitor is selected from the group consisting of 1,1,1-Trifluro-4-(4-hydroxyphenyl)butan-2-one (para), 1,1,1-Trifluro-4-(3-hydroxyphenyl)butan-2-one (meta), and 1,1,1-Trifluro-4-(2-hydroxyphenyl)butan-2-one (ortho).
19. The kit of claim 17 , wherein the antiviral drug is selected from the group consisting of Oseltamivir, Zanamivir, Lanamivir, and Peramivir.
20. The kit of claim 15 , further comprising reagents for an immunoassay.
21. The kit of claim 20 , wherein the immunoassay reagents are selected from the group consisting of an antibody specific to an influenza type A antigen, and an antibody specific to an influenza type B antigen.
22. A diagnostic system for detecting influenza virus comprising:
a cartridge for determining neuraminidase activity in a sample comprising a first buffer compatible with the neuraminidase determination;
a test for detecting influenza type A or type B in the sample, wherein the test is configured to process a sample comprising the first buffer into a sample comprising a second buffer, wherein the first buffer is incompatible with the test for detecting influenza type A or type B, and wherein the second buffer is compatible with the test for detecting influenza type A or type B; and
a first detector configured to measure a signal from the cartridge and/or the test.
23. The system of claim 22 , wherein the cartridge comprises neuraminidase activity assay reagents selected from the group consisting of luciferase, a conjugate of a N-acetylneuraminic acid and luciferin, a derivative of a N-acetylneuraminic acid and luciferin conjugate, an antiviral drug, and a reagent comprising a masked inhibitor compound for use in a dual enzyme influenza neuraminidase sensitivity assay.
24. The system of claim 23 , wherein the masked inhibitor is selected from the group consisting of 1,1,1-Trifluro-4-(4-hydroxyphenyl)butan-2-one (para), 1,1,1-Trifluro-4-(3-hydroxyphenyl)butan-2-one (meta), and 1,1,1-Trifluro-4-(2-hydroxyphenyl)butan-2-one (ortho).
25. The system of claim 22 , wherein the cartridge comprises a matrix chamber comprising a cartridge matrix comprising the first buffer for the determination, and a reagent chamber comprising reagents for the neuraminidase activity determination.
26. The system of claim 25 , wherein the cartridge matrix comprises a cross-linked polysaccharide.
27. The system of claim 25 , wherein the matrix chamber and reagent chamber are in fluid communication such that a sample applied to the matrix chamber flows from the matrix chamber to the reagent chamber.
28. The system of claim 22 , wherein the cartridge comprises a multiwell cartridge adapted to be read using a photomultiplier tube or micro-photomultiplier tube.
29. The system of claim 22 , wherein the test comprises an immunoassay.
30. The system of claim 22 , wherein the immunoassay is a sandwich assay.
31. The system of claim 22 , wherein the test comprises a test matrix comprising the second buffer for the detection.
32. The system of claim 31 , wherein the test matrix comprises nitrocellulose.
33. The system of claim 22 , wherein the second buffer for the detection is lyophilized or dried down.
34. The system of claim 22 , wherein the first detector is configured to measure a signal from the cartridge, and the system further comprises a second detector configured to measure a signal from the test.
35. The system of claim 34 , wherein the first detector comprises a photomultiplier tube or micro-photomultiplier tube, and the second detector comprises a luminometer.
36. A kit for detecting influenza virus comprising:
a cartridge for determining neuraminidase activity in a sample comprising a first buffer; and
a test for detecting influenza type A or type B in the sample, wherein the test is configured to process a sample comprising the first buffer into a sample comprising the second buffer, wherein the first buffer is incompatible with the test for detecting influenza type A or type B, and wherein the second buffer is compatible with the test for detecting influenza type A or type B.
37. The kit of claim 36 , further comprising reagents for a neuraminidase activity assay.
38. The kit of claim 37 , wherein the neuraminidase activity assay reagents are selected from the group consisting of luciferase, a conjugate of a N-acetylneuraminic acid and luciferin, a derivative of a N-acetylneuraminic acid and luciferin, a N-acetylneuraminic acid and a trifluoromethylketone, a derivative of a conjugate of a N-acetylneuraminic acid and a trifluoromethylketone, an antiviral drug, and a reagent comprising a masked inhibitor compound for use in a dual enzyme influenza neuraminidase sensitivity assay.
39. The kit of claim 38 , wherein the masked inhibitor is selected from the group consisting of 1,1,1-Trifluro-4-(4-hydroxyphenyl)butan-2-one (para), 1,1,1-Trifluro-4-(3-hydroxyphenyl)butan-2-one (meta), and 1,1,1-Trifluro-4-(2-hydroxyphenyl)butan-2-one (ortho).
40. The kit of claim 38 , wherein the antiviral drug is selected from the group consisting of Oseltamivir, Zanamivir, Lanamivir, and Peramivir.
41. The kit of claim 36 , further comprising reagents for an immunoassay.
42. The kit of claim 41 , wherein the immunoassay reagents are selected from the group consisting of an antibody specific to an influenza type A antigen, and an antibody specific to an influenza type B antigen.