FI112095B - Icke-radioaktivt lösningshybridiseringsbestämningsförfarande och -kit - Google Patents
Icke-radioaktivt lösningshybridiseringsbestämningsförfarande och -kit Download PDFInfo
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- FI112095B FI112095B FI942239A FI942239A FI112095B FI 112095 B FI112095 B FI 112095B FI 942239 A FI942239 A FI 942239A FI 942239 A FI942239 A FI 942239A FI 112095 B FI112095 B FI 112095B
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- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
- C12Q1/701—Specific hybridization probes
- C12Q1/706—Specific hybridization probes for hepatitis
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6813—Hybridisation assays
- C12Q1/6816—Hybridisation assays characterised by the detection means
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6865—Promoter-based amplification, e.g. nucleic acid sequence amplification [NASBA], self-sustained sequence replication [3SR] or transcription-based amplification system [TAS]
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/70—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving virus or bacteriophage
- C12Q1/701—Specific hybridization probes
- C12Q1/708—Specific hybridization probes for papilloma
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
- C12Q2525/30—Oligonucleotides characterised by their secondary structure
- C12Q2525/301—Hairpin oligonucleotides
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Wood Science & Technology (AREA)
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- Engineering & Computer Science (AREA)
- Immunology (AREA)
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- Genetics & Genomics (AREA)
- Virology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Communicable Diseases (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Claims (22)
1. Icke-radioaktivt lösningshybridiseringsbestäm-ningsförfarande för att detektera en mälnukleinsyrasekvens 5. ett biologiskt prov, kännetecknat av att det omfattar steg, där: a) en bas tillsätts provet till en slutlig kon-centration av 0,415 - 2 M vid en temperatur av 20 - 100 °C under en tid av 5 - 12 0 minuter för att hydrolysera pro- 10 vets RNA och denaturera malnukleinsyrasekvensen, b) nukleinsyrasekvensen i det behandlade provet hybridiseras till en komplementär nukleinsyrasond för att bilda en dubbelsträngad hybrid, c) hybriden fängas i en fast fas, till vilken en 15 antihybridantikropp eller ett funktionellt antihybridan- tikroppfragment har immobiliserats, varvid antikroppen eller antikroppfragmentet binds specifikt till en komponent i den dubbelsträngade hybriden, d) en eventuell icke-hybridiserad sond elimineras 20 genom nedbrytning med ett enzym och e) den bundna hybriden bestäms, varvid detekte- * ringen av den bundna hybriden kvantitativt anger närvaron : av malnukleinsyrasekvensen i provet.
2. Be stämningsf ör f arande enligt patentkrav 1, ·;· 25 kännetecknat av att mälnukleinsyran är DNA, som har : valts frän gruppen humant papillomavirus-DNA, hepatit B- DNA och Chlamydia-DNA.
3. Bestämningsförfarande enligt patentkrav 1, ,···, kännetecknat av att sonden är en RNA-sekvens, som är 3. komplementär till mäl-DNA.
• · : *· 4. Bestämningsförfarande enligt patentkrav l, • · » kännetecknat av att den dubbelsträngade hybriden är .:. en RNA/DNA-hybrid. • m » 112095 44
5. Bestämningsförfarande enligt patentkrav 1, kännetecknat av att det nedbrytande enzymet är ett RNAs.
6. Bestämningsförfarande enligt patentkrav 1, 5 kännetecknat av att sonden har en koncentration av 1 - 500 ng/ml.
7. Bestämningsförfarande enligt patentkrav 1, kännetecknat av att sonden har en koncentration av 20 - 200 ng/ml.
8. Bestämningsförfarande enligt patentkrav 1, kännetecknat av att sonden har en koncentration av cirka 75 ng/ml.
9. Bestämningsförfarande enligt patentkrav 1, kännetecknat av att basen är natriurnhydroxidsom, som 15 har en koncentration av 0,415 - 2,0 M och som inkuberas med provet vid en temperatur av 20 - 100 °C under en tid av 5 - 120 minuter.
10. Bestämningsförfarande enligt patentkrav 1, kännetecknat av att basen är natriurnhydroxidsom, som 2. har en koncentration av 0,415 - 0,8 M och som inkuberas med provet vid en temperatur av 60 - 70 °C under en tid av i » 30-60 minuter.
11. Bestämningsförfarande enligt patentkrav 1, kännetecknat av att basen är natriurnhydroxidsom, som • ·· 25 har en koncentration av 0,415 M och som inkuberas med pro- / vet vid en temperatur av cirka 65 °C under en tid av cirka : ; : 45 minuter.
12. Bestämningsförfarande enligt patentkrav 5, ;V. kännetecknat av att RNAset tillsätts i provet i en .···, 30 koncentration av 0,01 - 1 mg/ml och det inkuberas med pro- t t vet vid en temperatur av 4 - 45 °C under en tid av 5 minu-I ** ter-24 timmar.
13. Bestämningsförfarande enligt patentkrav 5, kännetecknat av att RNAset tillsätts i provet i en ttlt 35 koncentration av 0,05 - 0,5 mg/ml och det inkuberas med 112095 45 provet vid en temperatur av 20 - 30 °C under en tid av 10 - 60 minuter.
14. Bestämningsförfarande enligt patentkrav 5, kännetecknat av att RNAset tillsätts i provet i en 5 koncentration av 0,2 mg/ml och det inkuberas med provet vid rumstemperatur under en tid av cirka 30 minuter.
15. Bestämningsförfarande enligt patentkrav 1, kännetecknat av att däri upplöses sonden dessutom i en buffert, som äterställer provet till neutralt pH.
16. Bestämningsförfarande enligt patentkrav 15, kännetecknat av att bufferten innehäller 2-[bis (2-hydroxietyl)amino]etansulfonsyra och natriumacetat.
17. Lösningshybridiseringskit för detektering av en DNA-mälnukleinsyrasekvens för diagnostisering av genfel 15 eller mikrob- eller virusinfektioner i ett biologiskt prov med ett förfarande enligt patentkrav 1, kännetecknat av att det innehäller: a) ett provtransportunderlag för stabilisering av det biologiska provet, 2. b) en baslösning, vars baskoncentration är sädan att som upplöst provet erhälls en koncentration av 0,415 - 2 M, ., för hydrolysering av RNA i provet och för denaturering av *t ‘I mälnukleinsyrasekvensen däri, : c) en sond som är komplementär till den behandlade 25 mälnukleinsyrasekvensen för att bilda en dubbelsträngad nukleinsyrahybrid, V ·* d) en neutraliserande sonddiluent för att späda ut sonden och neutralisera basen, e) en fast fas, till vilken en antihybridantikropp 30 eller ett antihybridantikroppfragment har immobiliserats, ,,· varvid antikroppen är specifik för en komponent i hybriden ‘ ” som bildats i hybridiseringen av sonden och mälnukleinsy- • · '*··’ rasekvensen, ’·· f) ett nedbrytande enzym för att eliminera en • ;..j 35 eventuell icke-hybridiserad sond och 112095 46 g) ett medel för att detektera hybriden som bil-dats i hybridiseringen av sonden och mälnukleinsyrasekven-sen.
18. Kit enligt patentkrav 17, kännetecknat av 5 att mälnukleinsyran är DNA, som har valts frän gruppen hu- mant papillomavirus-DNA, hepatit B-virus-DNA och Chlamy-dia-DNA.
19. Kit enligt patentkrav 17, kännetecknat av att sonden är en RNA-sekvens, som är komplementär tili 10 mäl-DNA.
20. Kit enligt patentkrav 17, kännetecknat av att det nedbrytande enzymet är ett RNas.
21. Kit enligt patentkrav 17, kännetecknat av att basen är natriumhydroxidsom, som har en koncentration 15 av 0,415 - 2 M.
22. Kit enligt patentkrav 17, kännetecknat av att det nedbrytande enzymet och detekteringsmedlet ingär i samma reagens. * I · > I I * · · • · > I 1 > t » • · • · < ♦ 1 < · I « « » * »tl» » » » » · · I · I • » I ·
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US79258591A | 1991-11-14 | 1991-11-14 | |
US79258591 | 1991-11-14 | ||
US9209604 | 1992-11-12 | ||
PCT/US1992/009604 WO1993010263A1 (en) | 1991-11-14 | 1992-11-12 | Non-radioactive hybridization assay and kit |
Publications (3)
Publication Number | Publication Date |
---|---|
FI942239A0 FI942239A0 (sv) | 1994-05-13 |
FI942239A FI942239A (sv) | 1994-07-06 |
FI112095B true FI112095B (sv) | 2003-10-31 |
Family
ID=25157398
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
FI942239A FI112095B (sv) | 1991-11-14 | 1994-05-13 | Icke-radioaktivt lösningshybridiseringsbestämningsförfarande och -kit |
Country Status (11)
Country | Link |
---|---|
US (2) | US6228578B1 (sv) |
EP (1) | EP0667918B1 (sv) |
JP (1) | JP3091492B2 (sv) |
AT (1) | ATE189831T1 (sv) |
AU (1) | AU673813B2 (sv) |
DE (1) | DE69230693T2 (sv) |
DK (1) | DK0667918T3 (sv) |
ES (1) | ES2145750T3 (sv) |
FI (1) | FI112095B (sv) |
GR (1) | GR3033345T3 (sv) |
WO (1) | WO1993010263A1 (sv) |
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-
1992
- 1992-11-12 AU AU30689/92A patent/AU673813B2/en not_active Expired
- 1992-11-12 EP EP92924354A patent/EP0667918B1/en not_active Expired - Lifetime
- 1992-11-12 WO PCT/US1992/009604 patent/WO1993010263A1/en active IP Right Grant
- 1992-11-12 AT AT92924354T patent/ATE189831T1/de active
- 1992-11-12 DE DE69230693T patent/DE69230693T2/de not_active Expired - Lifetime
- 1992-11-12 DK DK92924354T patent/DK0667918T3/da active
- 1992-11-12 ES ES92924354T patent/ES2145750T3/es not_active Expired - Lifetime
- 1992-11-12 JP JP05509340A patent/JP3091492B2/ja not_active Expired - Lifetime
-
1994
- 1994-01-18 US US08/183,154 patent/US6228578B1/en not_active Expired - Lifetime
- 1994-05-13 FI FI942239A patent/FI112095B/sv not_active IP Right Cessation
-
2000
- 2000-04-26 GR GR20000401030T patent/GR3033345T3/el unknown
-
2001
- 2001-05-07 US US09/850,041 patent/US20020012936A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
JPH07505759A (ja) | 1995-06-29 |
ATE189831T1 (de) | 2000-03-15 |
AU673813B2 (en) | 1996-11-28 |
US20020012936A1 (en) | 2002-01-31 |
ES2145750T3 (es) | 2000-07-16 |
DE69230693D1 (de) | 2000-03-23 |
GR3033345T3 (en) | 2000-09-29 |
FI942239A (sv) | 1994-07-06 |
JP3091492B2 (ja) | 2000-09-25 |
FI942239A0 (sv) | 1994-05-13 |
WO1993010263A1 (en) | 1993-05-27 |
US6228578B1 (en) | 2001-05-08 |
DE69230693T2 (de) | 2000-06-15 |
AU3068992A (en) | 1993-06-15 |
DK0667918T3 (da) | 2000-06-05 |
EP0667918A1 (en) | 1995-08-23 |
EP0667918B1 (en) | 2000-02-16 |
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Legal Events
Date | Code | Title | Description |
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MA | Patent expired |