Claims (17)
1. Postupak dijagnosticiranja ili prognosticiranja Alzheimerove bolesti kod subjekta, naznačen time što se sastoji u najmanje jednom koraku otkrivanja prisutnosti polimorfije rs908832 gena ABCA2.1. A method of diagnosing or prognosticating Alzheimer's disease in a subject, characterized in that it consists in at least one step of detecting the presence of the polymorphism rs908832 of the ABCA2 gene.
2. Postupak u skladu s patentnim zahtjevom 1, naznačen time što se navedene korake otkrivanja provodi uz pomoć bioloških uzoraka.2. The method according to patent claim 1, characterized in that the specified detection steps are carried out with the help of biological samples.
3. Postupak dijagnosticiranja ili prognosticiranja Alzheimerove bolesti kod subjekta u skladu s patentnim zahtjevom 2, naznačen time što su navedeni biološki uzorci uzorci koji sadrže stanice s jezgrom.3. A method of diagnosing or prognosticating Alzheimer's disease in a subject according to claim 2, characterized in that said biological samples are samples containing nucleated cells.
4. Postupak dijagnosticiranja ili prognosticiranja Alzheimerove bolesti kod subjekta u skladu s patentnim zahtjevom 2 ili 3, naznačen time što su navedeni biološki uzorci uzorci krvi, uzorci sjemena ili uzorci dlake.4. A method of diagnosing or prognosticating Alzheimer's disease in a subject according to claim 2 or 3, characterized in that said biological samples are blood samples, semen samples or hair samples.
5. Postupak dijagnosticiranja ili prognosticiranja Alzheimerove bolesti kod subjekta u skladu s patentnim zahtjevom 1, naznačen time što navedene korake otkrivanja prisutnosti ili odsutnosti DNA koje nose navedenu polimorfiju se provodi uz pomoć tehnika lančane reakcije polimeraze (PCR), hibridizacije, bugačenja prema Southernu na membrani, digestije nukleazama, polimorfije duljine restrikcijskih fragmenata (RFLP) ili izravnog sekvenciranja, ili njihovom kombinacijom.5. A method of diagnosing or prognosticating Alzheimer's disease in a subject in accordance with claim 1, characterized in that the said steps of detecting the presence or absence of DNA carrying the said polymorphism are carried out with the help of polymerase chain reaction (PCR), hybridization, Southern blotting techniques on a membrane , nuclease digestion, restriction fragment length polymorphism (RFLP) or direct sequencing, or a combination thereof.
6. Postupak u skladu s patentnim zahtjevom 5, naznačen time što se sastoji u:
a. ekstrakciji DNA navedenog subjekta iz biološkog uzorka,
b. amplifikaciji navedene izolirane DNA uz pomoć početnica koje mogu amplificirati slijed koji odgovara polimorfiji rs908832 gena ABCA2,
c. određivanju prisutnosti najmanje jednog od alela polimorfije rs908832 gena ABCA2 u amplificiranoj DNA.6. The procedure according to patent claim 5, characterized in that it consists in:
a. DNA extraction of the said subject from a biological sample,
b. amplification of the mentioned isolated DNA with the help of primers that can amplify the sequence corresponding to the rs908832 polymorphism of the ABCA2 gene,
c. determining the presence of at least one allele of the polymorphism rs908832 of the ABCA2 gene in the amplified DNA.
7. Postupak u skladu s patentnim zahtjevom 6, naznačen time što se korak amplifikacije b) sastoji u lančanoj reakciji polimeraze (PCR).7. The method according to patent claim 6, characterized in that the amplification step b) consists in the polymerase chain reaction (PCR).
8. Postupak u skladu s patentnim zahtjevom 6, naznačen time što se amplificiranu DNA koja nosi manjinski alel "T" polimorfije rs908832 razlikuje od amplificirane DNA koja ne nosi taj alel tehnikom koja uključuje aktivnost 5' nukleaze DNA polimeraze I (TaqMan).8. The method according to patent claim 6, characterized in that the amplified DNA carrying the minority allele "T" of the rs908832 polymorphism differs from the amplified DNA not carrying that allele by a technique involving the activity of 5' nuclease DNA polymerase I (TaqMan).
9. Postupak u skladu s patentnim zahtjevom 6, naznačen time što se amplificiranu DNA koja nosi manjinski alel "T" polimorfije rs908832 razlikuje od amplificirane DNA koja ne nosi tu polimorfiju analizom polimorfije restrikcijskih fragmenata (RFLP).9. The method according to patent claim 6, characterized in that the amplified DNA carrying the minority allele "T" of the rs908832 polymorphism differs from the amplified DNA which does not carry this polymorphism by restriction fragment polymorphism (RFLP) analysis.
10. Postupak u skladu s patentnim zahtjevom 9, naznačen time što su restrikcijski fragmenti dobiveni digestijom amplificirane DNA restrikcijskim enzimom prije migracije u agarozni gel, bugačenja prema Southernu i hibridizacije.10. The method according to patent claim 9, characterized in that the restriction fragments are obtained by digesting the amplified DNA with a restriction enzyme before migration into an agarose gel, blotting according to Southern and hybridization.
11. Postupak u skladu s patentnim zahtjevom 1, naznačen time što se otkrivanje prisutnosti polimorfije rs908832 gena ABCA2 sastoji u određivanju nukleotida na položaju 2185 u transkriptu koji kodira protein ABCA2, gdje navedeni transkript sadrži slijed SEQ ID No.5.11. The method according to patent claim 1, characterized in that detecting the presence of polymorphism rs908832 of the ABCA2 gene consists in determining the nucleotide at position 2185 in the transcript encoding the ABCA2 protein, where said transcript contains the sequence of SEQ ID No.5.
12. Transgene životinje koje nisu ljudi, linije matičnih stanica i linije stanica koje se razlikuju od tih linija matičnih stanica, naznačene time što im je u genom ubačen najmanje jedan slijed egzogene genomske DNA koja nosi manjinski alel "T" polimorfije rs908832 gena ABCA2.12. Non-human transgenic animals, stem cell lines and cell lines that differ from these stem cell lines, characterized by the fact that at least one sequence of exogenous genomic DNA carrying the minority allele "T" of the rs908832 polymorphism of the ABCA2 gene has been inserted into their genome.
13. Transgene životinje u skladu s patentnim zahtjevom 12, naznačene time što su odabrane između sisavaca koji nisu ljudi.13. Transgenic animals according to claim 12, characterized in that they are selected from non-human mammals.
14. Postupak dobivanja transgenih životinja, linija matičnih stanica i linija stanica koje se razlikuju od tih linija matičnih stanica u skladu s bilo kojim od patentnih zahtjeva 12 i 13, naznačen time što se sastoji u generiranju transgenih životinja, s izuzetkom postupaka koji se sastoje u ili uključuju spolno križanje cijelog genoma tih životinja, linija matičnih stanica i linija stanica koje se razlikuju od tih linija matičnih stanica, koje sadrže manjinski alel polimorfije rs908832 ljudskog gena ABCA2.14. A process for obtaining transgenic animals, stem cell lines and cell lines that differ from these stem cell lines in accordance with any of claims 12 and 13, characterized in that it consists in the generation of transgenic animals, with the exception of the processes consisting in or involve whole-genome sexual crossing of those animals, stem cell lines, and cell lines different from those stem cell lines, which contain a minority allele of the rs908832 polymorphism of the human ABCA2 gene.
15. Upotreba transgenih životinja, linija matičnih stanica i linija stanica koje se razlikuju od tih linija matičnih stanica, u skladu s bilo kojim od patentnih zahtjeva 12 i 13, naznačena time što su transgene životinje i stanične linije namijenjene ispitivanju aktivnosti sredstava ili postupaka namijenjenih sprječavanju i/ili liječenju Alzheimerove bolesti.15. The use of transgenic animals, stem cell lines and cell lines different from these stem cell lines, in accordance with any of claims 12 and 13, characterized in that the transgenic animals and cell lines are intended for testing the activity of agents or procedures intended to prevent and/or treatment of Alzheimer's disease.
16. Stanica, naznačena time što je ekstrahirana iz transgenih životinja, kao što je opisano u bilo kojem od patentnih zahtjeva 12 i 13.16. A cell, characterized in that it is extracted from transgenic animals, as described in any of claims 12 and 13.
17. Upotreba stanice u skladu s patentnim zahtjevom 16, naznačena time što su stanice namijenjene ispitivanju spojeva namijenjenih liječenju Alzheimerove bolesti.
POPIS SLIJEDOVA
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[image] 17. Use of a cell according to claim 16, characterized in that the cells are intended for testing compounds intended for the treatment of Alzheimer's disease.
LIST OF SEQUENCES
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