DE164054T1 - Verfahren zum nachweis von polymorphischen restriktionsstellen und nukleinsauren sequenzen. - Google Patents

Verfahren zum nachweis von polymorphischen restriktionsstellen und nukleinsauren sequenzen.

Info

Publication number
DE164054T1
DE164054T1 DE198585106525T DE85106525T DE164054T1 DE 164054 T1 DE164054 T1 DE 164054T1 DE 198585106525 T DE198585106525 T DE 198585106525T DE 85106525 T DE85106525 T DE 85106525T DE 164054 T1 DE164054 T1 DE 164054T1
Authority
DE
Germany
Prior art keywords
restriction site
sample
restriction
nucleic acid
acid sequence
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
DE198585106525T
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English (en)
Inventor
Henry Anthony Oakland California 94602 Erlich
Randall Keichi Richmond California 94805 Saiki
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Novartis Vaccines and Diagnostics Inc
Original Assignee
Cetus Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cetus Corp filed Critical Cetus Corp
Publication of DE164054T1 publication Critical patent/DE164054T1/de
Pending legal-status Critical Current

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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
    • C12Q1/68—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
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6827—Hybridisation assays for detection of mutation or polymorphism
    • C12Q1/683—Hybridisation assays for detection of mutation or polymorphism involving restriction enzymes, e.g. restriction fragment length polymorphism [RFLP]
    • 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
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6832—Enhancement of hybridisation reaction
    • 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
    • C12Q1/68—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
    • C12Q1/686—Polymerase chain reaction [PCR]
    • 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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/803—Physical recovery methods, e.g. chromatography, grinding
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S435/00—Chemistry: molecular biology and microbiology
    • Y10S435/81—Packaged device or kit
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/807—Apparatus included in process claim, e.g. physical support structures
    • Y10S436/808—Automated or kit
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10—TECHNICAL SUBJECTS COVERED BY FORMER USPC
    • Y10S—TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y10S436/00—Chemistry: analytical and immunological testing
    • Y10S436/811—Test for named disease, body condition or organ function

Claims (8)

EP 85 10 6525.0 Cetus Corp. Deutsche Übersetzung der Patentansprüche nach Art. 67(1) EPÜ Patentansprüche:
1. Verfahren zum Nachweis der Anwesenheit oder Abwesenheit mindestens einer bestimmten Restriktionsstelle in einer bestimmten Nukleinsäure-Sequenz, gekennzeichnet durch die Stufen:
(a) Hybridisierung der Nukleinsäure-Sequenz mit einer Oligonukleotid-Probe für jede nachzuweisende Restriktionsstelle, wobei die Probe zu einem Bereich der Nukleinsäure-Sequenz komplementär ist, der die entsprechende Restrik-
tionsstelle der Probe überspannt und wobei die Probe an dem der jeweiligen Restriktionsstelle näheren Ende markiert ist;
(b) Verdauen der hybridisierten Nukleinsäure-Sequenz mit einer Restriktions-Endonuklease für jede Probe, wel-
ehe zur Spaltung der jeweiligen Probe an der nachzuweisenden Restriktionsstelle befähigt ist, wobei markierte und unmarkierte oligomere Fragmente erhalten werden, bei denen die Markierung die gleiche ist wie diejenige auf der Probe;
(c) Abtrennen aller markierten, gespalteten oligomeren Fragmente von markierten nichtgespalteten Oligomeren; und
(d) Nachweis der Gegenwart oder Abwesenheit von markierten oligomeren Fragmenten.
35
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Γ *■·'■·· ■-•-»2--·..·'..· 0164052
2. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß es ein Verfahren zum Nachweis der Gegenwart einer Restriktionsstelle ist, deren Bezug zu einer Infektionskrankheit bekannt ist.
3. Verfahren zum Nachweis der Gegenwart oder Abwesenheit von einem oder beiden Allelen einer polymorphen Restriktionsstelle in einer bestimmten Nukleinsäure-Sequenz, gekennzeichnet durch die Stufen:
(a) Hybridisierung der Nukleinsäure-Sequenz mit einer Oligonukleotid-Probe, welche komplementär ist zu einem Bereich in der Nukleinsäure-Sequenz, der eine nur in einem Alle! vorhandene erste Restriktionsstelle und eine beiden Allelen gemeinsame zweite Restriktionsstelle überspannt, und wobei die Probe an einem Ende markiert ist, das sich näher bei einer der genannten Restrictionsstellen befindet ;
(b) Verdauen der hybridisierten Nukleinsäure-Sequenz mit einer ersten Restriktionsendonuklease, welche zur Spaltung der Probe nur an der ersten Restriktionsstelle befähigt ist, wobei markierte und unmarkierte oligomere Fragmente erhalten werden, in denen die Markierung die gleiche wie diejenige an der Probe ist;
(c) Verdauen des Verdaus von Stufe (b) mit einer zweiten Restriktionsendonuklease, welche zur Spaltung der Probe nur an der zweiten Restriktionsstelle befähigt ist, wobei markierte und unmarkierte oligomere Fragmente erhalten werden, in denen die Markierung die gleiche wie diejenige der Probe ist;
(d) Abtrennung aller markierten, gespalteten oligomeren Fragmente von nichtgespalteten Oligomeren; und (e) Nachweis der Gegenwart oder Abwesenheit von markierten Fragmenten mit bestimmten Basenlängen, die mit der Spaltung durch die erste oder zweite Restriktionsendonuklease übereinstimmen.
4. Verfahren nach Anspruch 5? dadurch gekennzeichnet, daß die Krankheit Sichelzellanämie ist, die erste Sestriktionsstelle die in normalem ß-Globin vorhandene, in Sichelzellen-fi-Globin jedoch abwesende Ddel-Schnittstelle ist, und die zweite Restriktionsstelle, die sowohl in normalem als auch in Sichelzellen-ß-Globin vorhandene Hinf!-Schnittstelle ist.
5. "Verfahren nach einem der Ansprüche 1 bis 4, weiter da-
durch gekennzeichnet, daß nach der Stufe (a) und vor der Stufe (b) das Verfahren gekennzeichnet ist durch die Stufen:
(a1) Versetzen des Hybridisierungsgemisches von Stufe (a) mit einem unmarkierten blockierenden Oligomeren, das zu allen Proben mit Ausnahme von mindestens einem nicht richtig zusammenpassenden Basenpaar an jeder nachzuweisenden Restriktionsstelle komplementär ist; und (a") Hybridisieren des Hybridisierungsgemisches in Gegenwart des blockierenden Oligomeren.
6. Verfahren nach einem der Ansprüche 1 bis 5, dadurch gekennzeichnet, daß die Restriktionsstelle polymorph ist und die Hybridisierungsstufe in Gegenwart eines positiven Vergleichs-Oligomeren durchgeführt wird, das an einen Bereich der Nukleinsäure-Sequenz bindet, welcher eine nichtpolymorphe Restriktionsstelle für die Restriktionsendonuklease enthält.
7· Verfahren nach einem der Ansprüche 1 bis 6, dadurch gekennzeichnet, daß die Nukleinsäure-Sequenz eine DNA-Sequenz ist und die Probe ein Oligodeoxyribonukleotid ist.
8. Nachweissatz zum raschen Nachweis der Gegenwart oder Abwesenheit mindestens einer bestimmten Restriktionsstelle in einer bestimmten Nukleinsäure-Sequenz, gekennzeichnet durch:
L J
- ·ι+- 01
(a) eine Cligonukleotid-Probe für ,jede nachzuweisende Restriktionsstelle, wobei die Probe zu einem Bereich der Nukleinsäure-Sequenz komplementär ist, der die betreffende Restrik~ionsstelle der Probe überspannt, und wobei die Probe an dem der betreffenden Restriktionsstelle näheren Ende markiert ist; und
(b) eine Restriktionsendonuklease für jede Probe, die zur Spaltung der Probe an der Restriktionsstelie befähigt ist.
9· Nachweissatz nach Anspruch 8, weiter gekennzeichnet durch ein nichtmarkiertes blockierendes Oligomeres, das zu allen Proben mit Ausnahme von mindestens einem nichtpassenden Basenpaar an der Restriktionsstelle komplementär ist.
1C. Nachweissatz zum Nachweis der Gegenwart oder Abwesenheit von einem oder beiden Allelen einer poiymorphen Restriktionsstelle in einer bestimmten DNA-Sequenz, gekennzeichnet durch:
(a) eine Oligodeoxyribonukleotid-Probe, welche zu einem Bereich in der DNA-Sequenz komplementär ist, der eine erste, nur in einem Allel vorhandene Restriktionsstelle und eine zweite, beiden Allelen gemeinsame Restriktionsstellle überspannt und wobei die Probe an dem zu einer der Restriktionsstellen näheren Ende markiert ist; ("b) eine erste Restriktionsendonuklease, die zur Spaltung der Probe nur an der ersten Restriktionsstelle befähigt ist; und
(c) eine zweite Restriktionsendonuklease, die zur Spaltung der Probe nur an der zweiten Restriktionsstelle befähigt ist.
DE198585106525T 1984-05-29 1985-05-28 Verfahren zum nachweis von polymorphischen restriktionsstellen und nukleinsauren sequenzen. Pending DE164054T1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US61495784A 1984-05-29 1984-05-29
US06/716,982 US4683194A (en) 1984-05-29 1985-03-28 Method for detection of polymorphic restriction sites and nucleic acid sequences

Publications (1)

Publication Number Publication Date
DE164054T1 true DE164054T1 (de) 1986-04-30

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DE8585106525T Expired DE3572693D1 (en) 1984-05-29 1985-05-28 Method for detection of polymorphic restriction sites and nucleic acid sequences
DE198585106525T Pending DE164054T1 (de) 1984-05-29 1985-05-28 Verfahren zum nachweis von polymorphischen restriktionsstellen und nukleinsauren sequenzen.

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DE8585106525T Expired DE3572693D1 (en) 1984-05-29 1985-05-28 Method for detection of polymorphic restriction sites and nucleic acid sequences

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US (1) US4683194A (de)
EP (1) EP0164054B1 (de)
JP (1) JPS6119500A (de)
AU (1) AU587189B2 (de)
CA (1) CA1242627A (de)
DE (2) DE3572693D1 (de)
DK (1) DK240285A (de)
ES (1) ES8608584A1 (de)
IL (1) IL75327A (de)

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US4683194A (en) 1987-07-28
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