DE79139T1 - Methode und reagenskombination zur indentifikation von mikroorganismen und die anwendung der sandwichhybridisierung von nukleinsaeuren hierfuer. - Google Patents

Methode und reagenskombination zur indentifikation von mikroorganismen und die anwendung der sandwichhybridisierung von nukleinsaeuren hierfuer.

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Publication number
DE79139T1
DE79139T1 DE198282305489T DE82305489T DE79139T1 DE 79139 T1 DE79139 T1 DE 79139T1 DE 198282305489 T DE198282305489 T DE 198282305489T DE 82305489 T DE82305489 T DE 82305489T DE 79139 T1 DE79139 T1 DE 79139T1
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Prior art keywords
nucleic acid
microorganisms
pair
reagents
complementary
Prior art date
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Pending
Application number
DE198282305489T
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English (en)
Inventor
Tuula Marjut Fi-02660 Espoo 66 Ranki
Hans Erik Fi-02940 Espoo 94 Soederlund
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ORION CORP Ltd 00510 HELSINKI FI
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ORION CORP Ltd 00510 HELSINKI FI
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Application filed by ORION CORP Ltd 00510 HELSINKI FI filed Critical ORION CORP Ltd 00510 HELSINKI FI
Publication of DE79139T1 publication Critical patent/DE79139T1/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
    • C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
    • C12Q1/04—Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor
    • 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
    • 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
    • C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
    • 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
    • C12Q1/6888—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
    • 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
    • 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
    • Y10S436/804—Radioisotope, e.g. radioimmunoassay
    • 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
    • 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
    • Y10S436/823—Immunogenic carrier or carrier per se

Claims (12)

Ansprüche
1. Anwendung der Sandwich-Hybridisierungstechnik fUr Nukleinsäuren, wobei diese Technik darin besteht, dass man einsträngige NukLeinsäure aus einem Mikroorganismus mit einem Paar verschiedener Nukleinsäurereagenzien in Berührung bringt, wobei beide Reagenzien des Paars einsträngig und mit der vom Mikroorganismus abgeleiteten NukLeinsäure komplementär sind und ein Glied des Paars ein an einen festen Träger gebundenes NukIeinsäurefragment ist, während das andere ein mit einem Indikator markiertes Nukleinsäurefragment darstellt, wodurch sich ein an den festen Träger gebundenes markiertes Hybrid bildet, auf die Identifizierung eines in einer Probe vorliegenden Mikroorganismus bzw. einer Gruppe von Mikroorganismen, wobei das Paar NukLeinsäurereagenzien entsprechend dem Mikroorganismus oder der Gruppe von Mikroorganismen, deren Gegenwart in der Probe zu erwarten ist, ausgewählt und die Richtigkeit der Identifizierung durch Erfassung des Ausmasses der Bildung eines an den festen Träger gebundenen markierten Hybrids nachgewiesen wird.
2. Anwendung der Sandwich-Hybridisierungstechnik für Nukleinsäure nach Anspruch 1, dadurch gekennzeichnet, dass man mehrere NukLeinsäurereagenzienpaare verwendet, wobei jedes Paar mit von einem verschiedenen Mikroorganismus bzw. einer Gruppe von Mikroorganismen, wovon irgendeiner in der Probe zu erwarten ist, abgeleiteten Nukleinsäure komplementär ist.
3. Anwendung der Sandwich-Hybridisierungstechnik für Nukleinsäure nach Anspruch 2, dadurch gekennzeichnet, dass mehrere verschiedene Mikroorganismen bzw. Gruppen von Mikroorganismen in der Probe vorliegen und in dersel-
0079 Ί
ben ungeteiLten Probe durch Anwendung einer Reihe verschiedener Paare von NukIeinsäurereagenzien, die mit Nukleinsäuren aus den verschiedenen Mikroorganismen bzw. Gruppen von Mikroorganismen, deren Gegenwart in der Probe zu erwarten ist, komplementär sind /nachgewiesen werden.
4. Methode zu Identifizierung von in einer Probe vorliegenden Mikroorganismen bzw. Gruppen von Mikroorganismen durch Sandwich-Hybridisierung von Nukleinsäuren unter Verwendung eines Paars völlig verschiedener Nukleinsäurereagenzien, die beide mit Nukleinsäure aus demselben bekannten Mikroorganismus bzw. Gruppe vr>n Mikroorganismen komplementär und damit hybridisierbar sind, wobei ein Glied des Paars von NukLeinsäurereagenzien ein an einen festen Träger gebundenes einsträngiges NukLeinsäurefragment und das andere ein mit einem Indikator markiertes einsträngiges Nukleinsäurefragment ist, wodurch sich bei gelungener Hybridisierung ein an den festen Träger gebundenes markiertes Hybrid biLdet, dadurch gekennzeichnet, dass mehrere in einer einzigen ungeteiLten Probe vorhandene Mikroorganismen bzw. Gruppen von Mikroorganismen identifiziert werden, indem man Nukleinsäuren aus den Mikroorganismen in der Probe, nachdem sie einsträngig gemacht wurden, mit mehreren der Paare völlig verschiedener NukLeinsäurereagentien in Berührung bringt, wobei die Reagenzien in unterschiedlichen Paaren mit Nukleinsäure aus verschiedenen bekannten Mikroorganismen bzw. Gruppen von Mikroorganismen komplementär und damit hybridisierbar sind, und die Bildung oder Nichtbildung an einen festen Träger gebundener markierter Hybride fUr jedes der Paare von NukLeinsäurereagenzien nachweist.
5. Reagenzkombination zur Verwendung nach Anspruch 2 oder 3 bzw. fUr die Methode nach Anspruch 4, gekennzeichnet durch die Gegenwart von mehreren Paaren verschiedener NukLeinsäurereagenzien, wobei die beiden Reagenzien desselben Paars jeweils mit der NukLeinsäure aus demselben Mikroorganismus bzw. Gruppe von Mikroorganismen komplementär und damit hybridisierbar sind und die Reagenzien unterschiedlicher Paare mit NukLeinsäure aus unter-
schied I ichen Mikroorganismen bzw. Gruppen von Mikroorganismen komplementär und damit hybridisierbar sind, wobei ein Reagenz, aus jedem Paar ein an einen festen Träger gebundenes einsträngiges NukLeinsäurefragment und das andere Reagenz in jedem Paar ein mit einem Indikator markiertes einstrangiges Nuk I einsäurefragment ist. 6. Reagenzkombination nach Anspruch 5, dadurch gekennzeichnet, dass die beiden NukI einsäurereagenzien im selben Paar jeweils entweder direkt aus dem Genom des Mikroorganismus, mit dem sie komplementär sind, oder durch eine DNA-Rekombinationstechnik und nachherige Einsträngigmachung hergestellte Fragmente sind sowie eines der Reagenzien an einen festen Träger gebunden und das andere mit einem Indikator markiert ist.
7. Reagenzkombination nach Anspruch 5 oder 6, dadurch gekennzeichnet, dass eines oder mehrere der Paare Nukleinsäure reagenz i en mit Nukleinsäure aus einem fUr eine Infektion der Atemwege oder Durchfall verantwortlichen B a k terium oder Virus, einem fUr eine Geschlechtskrankheit verantwortlichen Bakterium, Virus, einer Hefe oder einem Protozoon oder einem für Sepsis oder unzureichende Lebensmittelhygiene verantwortlichen Bakterium komplementär und damit hybridisierbar sind.
8. Reagenzkombination nach Anspruch 5 oder 6, dadurch 5 gekennzeichnet, dass eines der Paare Nukleinsäurereagenzien mit Nukleinsäure aus Adenovirus komplementär und damit hybridisierbar ist, wobei dieses Paar aus dem an einen festen Träger gebundenen Adenovirusrekombinationsp lasmid Ad_DpBR322 und dem mit einem Indikator markierten Adenovirus-Ad -BamHI-C-Fragment besteht.
9. Reagenzkombination nach einem der Ansprüche 5 bis 8, dadurch gekennzeichnet, dass eines der Paare Nukleinsäurereagenzien mit Nukleinsäure aus Semliki-Forest-Virus komplementär und damit hybridisierbar ist, wobei dieses Paar aus dem an einen festen Träger gebundenen EcoRI-XhoI-Fragment A des pKTH312-Plasmids und dem mit einem Indikator markierten EcoRI-Xhol-Fragment B des pKTH312-Plasmids besteht.
10." Reagenzkombination nach einem der Ansprüche 5 bis
00791339, dadurch gekennzeichnet, dass eines der Paare Nukleinsäurereagenzoen mit Nukleinsäure aus SV40-Virus komplementär und damit hybridisierbar ist, wobei dieses Paar aus dem an einen festen Träger gebundenen PstI-B-Fragment des SV40-Virus und dem mit einem Indikator markierten Pst I-A-Fragment des SV40-Virus besteht.
11. Reagenzkombination nach einem der Ansprüche 5 bis
10, dadurch gekennzeichnet, dass eines der Paare Nukleinsäure reagen zi en mit NukLeinsäure aus Bacillus amylolique fax Jens komplementär und damit hybridisierbar ist, wobei dieses Paar aus dem an einen festen Träger gebundenen EcoRl-BamHI-Fragment des cc-Amyla Segens aus Bacillus a m y IoIiquefa-eiens-Plasmid pKTHiO und dem mit einem Indikator markierten Clal-EcoRI-Fragment des α-Amy lasegens aus Ba-5 eil I us amy Io I iquefaciens-Plasmid ρ K T H1 0 besteht.
12. Reagenzkombination nach Anspruch 5, dadurch gekennzeichnet, dass eines der Paare NukIeinsäurereagenzien mit Nukleinsäure aus E. coli komplementär und damit hybridisierbar ist, wobei dieses Paar aus dem an einen festen Träger gebundenen Plasmid pKTH45 und dem mit einem
Indikator markierten Rekombinationsphagen mKTHi207 besteht.
DE198282305489T 1981-10-16 1982-10-15 Methode und reagenskombination zur indentifikation von mikroorganismen und die anwendung der sandwichhybridisierung von nukleinsaeuren hierfuer. Pending DE79139T1 (de)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
FI813251A FI63596C (fi) 1981-10-16 1981-10-16 Mikrobdiagnostiskt foerfarande som grundar sig pao skiktshybridisering av nukleinsyror och vid foerfarandet anvaenda kombinationer av reagenser

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DE79139T1 true DE79139T1 (de) 1983-11-24

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DE8282305489T Expired DE3277917D1 (en) 1981-10-16 1982-10-15 A method and reagent combination for the identification of microorganisms and the use of sandwich hybridization of nucleic acids therefor
DE198282305489T Pending DE79139T1 (de) 1981-10-16 1982-10-15 Methode und reagenskombination zur indentifikation von mikroorganismen und die anwendung der sandwichhybridisierung von nukleinsaeuren hierfuer.

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DE8282305489T Expired DE3277917D1 (en) 1981-10-16 1982-10-15 A method and reagent combination for the identification of microorganisms and the use of sandwich hybridization of nucleic acids therefor

Country Status (13)

Country Link
US (2) US4486539A (de)
EP (2) EP0098267A1 (de)
JP (2) JPH0632637B1 (de)
AT (1) ATE31735T1 (de)
AU (1) AU548854B2 (de)
CA (1) CA1192120A (de)
DE (2) DE3277917D1 (de)
DK (1) DK173744B1 (de)
FI (1) FI63596C (de)
HU (1) HU196242B (de)
RO (1) RO86356B (de)
SU (1) SU1386031A3 (de)
WO (1) WO1983001459A1 (de)

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AU8957582A (en) 1983-05-05
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DK275183A (da) 1983-06-15
FI63596B (fi) 1983-03-31
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HU196242B (en) 1988-10-28
US4563419A (en) 1986-01-07
RO86356A (ro) 1985-03-15
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