1. An apparatus for regulating the delivery of a liquid from a bottle, comprising:
a housing covering an opening of the bottle;
a flow regulating cartridge, said cartridge being removably inserted within said housing and extending into an opening of the container, said regulating cartridge making contact with the opening to removably affix both the regulating cartridge and the housing to cover the opening;
an air vent, operatively associated with said regulating cartridge, said air vent equalizing air pressure within the bottle as liquid is poured through said regulating cartridge;
a percolation labyrinth, associated with said regulating cartridge, said labyrinth causing a turbulent flow of the liquid poured from the container through the housing, wherein said labyrinth a rolled spiral coil; and
a deformable cap for sealing the bottle by attachment to a rim thereon.
2. The apparatus according to claim 1 wherein the housing further includes a filter screen therewithin through which the liquid passes while being poured.
3. The apparatus according to claim 1 wherein said bottle is a wine bottle and the liquid is wine.
4. The apparatus according to claim 1 wherein said labyrinth consists essentially of #20 mesh of stainless steel rolled into the spiral coil.
5. The apparatus of claim 1, wherein the housing includes an outer tapered surface and where a surface of a lower portion of said cap are similarly tapered to match the outer tapered surface.
6. The apparatus of claim 5, wherein said cap further includes a tab on an upper surface thereof.
7. The apparatus of claim 1, wherein said flow regulating cartridge produces a linear flow rate of between about 1.0 and 2.0 ounces per second of liquid poured therethrough, and concurrently aerates the liquid.
8. The apparatus of claim 1, wherein said flow regulating cartridge produces a linear flow rate of between about 1.2 and 1.8 ounces per second of liquid poured therethrough.
9. The apparatus of claim 8, wherein said flow regulating cartridge concurrently aerates the liquid flowing therethrough.
10. The apparatus according to claim 1 wherein at least said housing is formed of stainless steel.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A method for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6 comprising the step of designing a set of primers per genotype in the NS34A genomic region, wherein said set of primers for amplifying said HCV NS34A nucleic acid is characterized in that at least one of the primers is an antisense primer that is capable of annealing to the NS34A genomic region spanning nucleotides 5622 to 5656, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
2. The method of claim 1 for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6 wherein said set of primers is further characterized in that it comprises a sense primer that is capable of annealing to the NS34A genomic region spanning nucleotides 3269 to 3295, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
3. The method of claim 1 for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6 wherein said set of primers is further characterized in that it comprises a sense primer that is capable of annealing to the NS34A genomic region spanning nucleotides 3995 to 4014, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
4. The method of claim 1 for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6 wherein said set of primers is further characterized in that it comprises a sense primer that is capable of annealing to the NS34A genomic region spanning nucleotides 4678 to 4736, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
5. A method for amplification of an HCV NS3 nucleic acid of any one of HCV genotypes 5 to 6 comprising the step of designing a set of primers per genotype in the NS3 genomic region, wherein said set of primers for amplifying said HCV NS3 nucleic acid is characterized in that at least one of the primers is a sense primer that is capable of annealing to the NS3 genomic region spanning nucleotides 3269 to 3295, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
6. The method of claim 5 for amplification of an HCV NS3 nucleic acid of any one of HCV genotypes 5 to 6, wherein said set of primers is further characterized in that it comprises an antisense primer that is capable of annealing to the NS3 genomic region spanning nucleotides 4989 to 5011, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
7. The method of claim 5 for amplification of an HCV NS3 nucleic acid of any one of HCV genotypes 5 to 6, wherein said set of primers is further characterized in that it comprises an antisense primer that is capable of annealing to the NS3 genomic region spanning nucleotides 4290 to 4351, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
8. A method for amplification of an HCV NS3 nucleic acid of HCV genotype 1 comprising the step of designing a set of primers in the NS3 genomic region, wherein said set of primers for amplifying said HCV NS3 nucleic acid is characterized in that at least one of the primers is an antisense primer that is capable of annealing to the NS3 genomic region spanning nucleotides 4917 to 4935, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
9. The method of claim 8 for amplification of an HCV NS3 nucleic acid of HCV genotype 1 wherein said set of primers is further characterized in that it comprises a sense primer that is capable of annealing to the NS3 genomic region spanning nucleotides 4137 to 4158, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
10. A method for amplification of an HCV NS34A or NS3 nucleic acid of any one of HCV genotypes 5 to 6 or HCV genotype 1 comprising the step of designing a set of primers per genotype in the NS34A or NS3 genomic region, wherein said set of primers for amplifying said HCV NS34A or NS3 nucleic acid is characterized in that one of the primers is an antisense primer chosen from the group consisting of
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 5622 to 5656,
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4989 to 5011,
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4290 to 4351,
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4917 to 4935,
and that the other primer is a sense primer chosen from the group consisting of
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 3269 to 3295,
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 3995 to 4014,
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4678 to 4736,
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4137 to 4158,
and wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
11. A method for determining the sequence of an HCV NS34A or NS3 nucleic acid of any one of HCV genotypes 5 to 6 or HCV genotype 1 wherein said NS34A or NS3 nucleic acid is amplified according to the method of any of claims 1 to 10 prior to or concurrent with determining said sequence.
12. A method for evaluating the sequence variations in an HCV NS34A or NS3 nucleic acid of any of HCV genotypes 5 to 6 or HCV genotype 1 wherein the sequence of said NS34A or NS3 nucleic acid is determined according to the method of claim 11 prior to evaluating the sequence variations.
13. The method of claim 12 wherein said sequence variations are mutations that are induced by HCV antiviral agents.
14. The method of claim 12 or 13 wherein said sequence variations arc mutations causing HCV drug resistance.
15. The method of claim 12 wherein said sequence variations are genotype-specific nucleotide variations.
16. The method of any of claims 1 to 15 wherein said amplification is a one-step RT-PCR.
17. The method of any of claims 1 to 10 which is part of a multiplex amplification method.
18. The method of any of claims 1 to 17 wherein the HCV NS34A or NS3 nucleic acid is present in a biological sample.
19. A recombinant vector comprising an NS34A or NS3 fragment amplified according to the method of any one of claims 1 to 10.
20. A set of primers for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6, said set of primers characterized in that at least one of the primers is an antisense primer that is capable of annealing to the HCV NS34A genomic region spanning nucleotides 5622 to 5656, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
21. The set of primers according to claim 20 for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6, said set of primers further characterized in that it comprises a sense primer that is capable of annealing to the HCV NS34A genomic region spanning nucleotides 3269 to 3295, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
22. The set of primers according to claim 20 for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6, said set of primers further characterized in that it comprises a sense primer that is capable of annealing to the HCV NS34A genomic region spanning nucleotides 3995 to 4014, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
23. The set of primers according to claim 20 for amplification of an HCV NS34A nucleic acid of any one of HCV genotypes 5 to 6, said set of primers further characterized in that it comprises a sense primer that is capable of annealing to the HCV NS34A genomic region spanning nucleotides 4678 to 4736, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
24. A set of primers for amplification of an HCV NS3 nucleic acid of any one of HCV genotypes 5 to 6, said set of primers characterized in that at least one of the primers is a sense primer that is capable of annealing to the HCV NS3 genomic region spanning nucleotides 3269 to 3295, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
25. The set of primers according to claim 24 for amplification of an HCV NS3 nucleic acid of any one of HCV genotypes 5 to 6, said set of primers further characterized in that it comprises an antisense primer that is capable of annealing to the HCV NS3 genomic region spanning nucleotides 4989 to 5011, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
26. The set of primers according to claim 24 for amplification of an HCV NS3 nucleic acid of any one of HCV genotypes 5 to 6, said set of primers further characterized in that it comprises an antisense primer that is capable of annealing to the HCV NS3 genomic region spanning nucleotides 4290 to 4351, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
27. A set of primers for amplification of an HCV NS3 nucleic acid of HCV genotype 1, said set of primers characterized in that at least one of the primers is an antisense primer that is capable of annealing to the HCV NS3 genomic region spanning nucleotides 4917 to 4935, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
28. The set of primers according to claim 27 for amplification of an HCV NS3 nucleic acid of HCV genotype 1, said set of primers further characterized in that it comprises a sense primer that is capable of annealing to the HCV NS3 genomic region spanning nucleotides 4137 to 4158, wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
29. A set of primers for amplification of an HCV NS34A or NS3 nucleic acid of any one of HCV genotypes 5 to 6 or HCV genotype 1, said set of primers characterized in that one of the primers is an antisense primer chosen from the group consisting of
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 5622 to 5656,
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4989 to 5011,
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4290 to 4351,
an antisense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4917 to 4935,
and that the other primer is a sense primer chosen from the group consisting of
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 3269 to 3295,
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 3995 to 4014,
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4678 to 4736,
a sense primer capable of annealing to the NS34A or NS3 genomic region spanning nucleotides 4137 to 4158,
and wherein the nucleotide positions correspond with the nucleotide numbering for HCV strain HCV-H.
30. Use of a set of primers according to claims 20 to 29 for the manufacture of a kit for amplification of an HCV NS34A or NS3 nucleic acid of any one of HCV genotypes 5 to 6 or HCV genotype 1.
31. A kit for amplification of an HCV NS34A or NS3 nucleic acid of any one of HCV genotypes 5 to 6 or HCV genotype 1, said kit comprising a set of primers according to claims 20 to 29.
32. The kit according to claim 31 which is a kit for multiplex amplification.