1. A video coding method applied to an original video sequence in which the successive frames or video object planes (VOPs) include one or several arbitrarily shaped video objects (VOs) defined in each VOP by their texture and motion components and an additional shape component, said method comprising the following steps
(a) a non object-oriented coding step, applied to a small number of frames of the video sequence ;
(b) an object-oriented coding step, applied to all the frames of the sequence that follow said small number of frames;
(c) a sequencing step, provided for controlling that said non object-oriented and object-oriented coding steps are respectively applied to the appropriate frames, in order to generate a coded bitstream including non object-oriented coded data corresponding to said small number of frames followed by object-oriented coded data corresponding to said following frames.
2. A coding method according to claim 1, in which said number of frames is equal to two.
3. A coding method according to claim 1, wherein said coded bitstream also includes an information about the number of regions of interest in the original video sequence.
4. A coding method according to claim 3, wherein said information about the number of regions of interest is given at the picture level.
5. A video decoding method applied to a coded bitstream corresponding to an original video sequence in which the successive frames or video object planes (VOPs) include one or several arbitrarily shaped video objects (VOs) defined in each VOP by their texture and motion components and an additional shape component and have been coded by means of a video coding method comprising the following steps:
(a) a non object-oriented coding step, applied to a small number of frames of the video sequence ;
(b) an object-oriented coding step, applied to all the frames of the sequence that follow said small number of frames;
(c) a sequencing step, provided for controlling that said non object-oriented and object-oriented coding steps are respectively applied to the appropriate frames, in order to generate a coded bitstream including non object-oriented coded data corresponding to said small number of frames followed by object-oriented coded data corresponding to said following frames;
said decoding method itself comprising the following steps
(1) a first decoding step, applied to said non object-oriented coded data of the coded bitstream that correspond to said small number of frames of the original video sequence;
(2) a spatio-temporal segmentation step applied to said non object-oriented coded data of the coded bitstream that correspond to said small number of frames and provided for reconstructing the missing shape component of the VOs;
(3) a second decoding step, applied to said object-oriented coded data of the coded bitstream that correspond to said following frames;
(4) a sequencing step, provided for controlling that said decoding and segmentation steps are respectively applied to the appropriate frames.
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. Primers for amplifying an HLA gene.
2. The primers of claim 1, wherein the HLA gene is HLA A, B or DRB1 or an exon therein.
3. The primers of claim 2, wherein the HLA-A primers have sequences shown in SEQ ID NOS: 14-15, the HLA-A exon 2 primers have the sequences shown in SEQ ID NOS: 20-21 and the HLA-A exon 3 primers have the sequences shown in SEQ ID NOS: 22-26.
4. The primers of claim 2, wherein the HLA-B primers have sequences shown in SEQ ID NOS: 16-19, the HLA-B exon 2 primers have the sequences shown in SEQ ID NOS: 27-28 and the HLA-B exon 3 primers have the sequences shown in SEQ ID NOS: 29-31.
5. The primers of claim 2, wherein the HLA-DRB1 primers have sequences shown in SEQ ID NOS: 32-37 and the HLA-B exon 2 primers have the sequences shown in SEQ ID NOS: 38-47.
6. Hybridization probes for detecting single nucleotide polymorphisms (SNPs) in an HLA gene, comprising:
about a 9-15mer oligonucleotide complementary to a region containing the SNP; and
5\u2032 and 3\u2032 flanking sequences.
7. The hybridization probes of claim 6, wherein the flanking sequences are oligo-thymidines or an oligo-thymidine-like polyanionic polymer.
8. The hybridization probes of claim 6, wherein the SNPs are located in HLA-A exon 2 or exon 3, HLA-B exon 2 or exon 3 or HLA-DRB1 exon 2.
9. The hybridization probes of claim 8, wherein the HLA-A exon 2 probes has the sequence shown in SEQ ID NO: 49 and the HLA-A exon 3 probes have the sequences shown in SEQ ID NOS: 101-153.
10. The hybridization probes of claim 8, wherein the HLA-B exon 2 probes have the sequences shown in SEQ ID NOS: 154-237 and the HLA-B exon 3 probes have the sequences shown in SEQ ID NOS: 238-239.
11. A microarray device for allelotyping an HLA gene, comprising:
a substrate having a cationic surface; and
a monolayer comprising one or more of the hybridization probes of claim 6 adsorbed thereto.
12. The microarray device of claim 11, wherein the cationic surface comprises an aminosilane, a quanidinium, tin oxide, aluminum oxide or zirconium oxide or other equivalently charged moiety.
13. The microarray device of claim 11, wherein the substrate is glass, plastic or metal.
14. The microarray device of claim 11, further comprising:
an oligo-thymidine co-absorbed with the hybridization probes.
15. The microarray device of claim 14, wherein the oligo-thymidine has about 20 to about 40 thymidine.
16. The microarray device of claim 14, further comprising:
a fluorescent dye linked to the oligo-thymidine
17. The microarray device of claim 11, further comprising:
a capping agent.
18. A kit for population-scale HLA genotyping, comprising:
gene-specific primers for amplifying an HLA gene; and
the microarray device of claim 11.
19. The kit of claim 18, wherein the primers have the sequences shown in SEQ ID NOS: 14-47.
20. The kit of claim 18, further comprising:
buffers and polymerases for a PCR reaction or a fluorescent dye or a combination thereof.
21. A system for real-time high throughput population-scale HLA allelotyping in a field environment, comprising:
the microarray device of claim 11;
means for collecting and purifying DNA samples from individuals comprising a population;
means for generating by PCR cRNA target amplicons of one or more HLA genes of interest from the collected DNA; and
means for assigning an HLA allelotype to each individual HLA gene of interest;
wherein individual means and devices comprising said system are portable and operable in real time within the field environment.
22. The system of claim 21, wherein the HLA gene is HLA-A, HLA-B or HLA-DRB1.
23. The system of claim 21, wherein the means for collecting DNA samples comprises:
a container suitable to receive a buccal wash sample, a buccal swab sample or a blood sample collected from the individuals.
24. The system of claim 21, wherein the means for generating target amplicons comprises HLA gene-specific primers for amplifying the HLA gene of interest.
25. The system of claim 24, wherein the gene-specific primers have sequences shown in SEQ ID NOS: 14-47.
26. The system of claim 21, wherein said means for assigning an HLA-allelotype to each individual comprises:
an imaging device adapted to detect hybridization patterns formed on the microarray device after hybridization of the target to the hybridization probes adsorbed thereto; and
pattern recognition software comprising a set of algorithms adapted to recognize the imaged hybridization patterns as HLA allelotypes.
27. The system of claim 26, wherein the hybridization probes have sequences shown in SEQ ID NOS: 48-289.
28. The system of claim 21, wherein real time high throughput genotyping is about 200 to about 300 HLA allelotypes per hour per system operated.
29. A method for real time population-scale HLA allelotyping in a field environment, comprising:
collecting DNA from one or members of the population;
purifying the DNA for analysis;
generating a target amplicon from an HLA gene of interest comprising the DNA using gene specific primers;
contacting the hybridization probes comprising the microarray of claim 11 with the target; and
imaging the hybridization pattern formed after the contact wherein each HLA allelotype has a pattern associated therewith.
30. The method of claim 29, further comprising storing the collected DNA.
31. The method of claim 29, wherein the DNA is collected from blood, with a buccal wash or with a buccal swab.
32. The method of claim 29, wherein the gene-specific primers have sequences shown in SEQ ID NOS: 14-47 and the hybridization probes have sequences shown in SEQ ID NOS: 48-289.
33. The method of claim 29, further comprising one or both of:
assessing a risk of infection by a biological agent or weapon for each individual based on the assigned allelotype; or
assessing a response to a particular vaccine against the biological agent or weapon by each individual; or
34. The method of claim 29, wherein the allelotype assigned to each individual of the population comprises a means of identification thereof.