DK200401109A - Kit for Multiplication and Detection of Nucleic Acid Sequences - Google Patents

Kit for Multiplication and Detection of Nucleic Acid Sequences Download PDF

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Publication number
DK200401109A
DK200401109A DK200401109A DKPA200401109A DK200401109A DK 200401109 A DK200401109 A DK 200401109A DK 200401109 A DK200401109 A DK 200401109A DK PA200401109 A DKPA200401109 A DK PA200401109A DK 200401109 A DK200401109 A DK 200401109A
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DK
Denmark
Prior art keywords
nucleic acid
kit
acid sequences
multiplication
detection
Prior art date
Application number
DK200401109A
Other languages
Danish (da)
Inventor
Scharf Stephen Joel
Horn Glenn Thoman
Ehrlich Henry Anthony
Saiki Randall Keichi
Arnheim Norman
Mullis Kary Banks
Original Assignee
Hoffmann La Roche
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
Priority claimed from US06/791,308 external-priority patent/US4683202A/en
Priority claimed from US06/828,144 external-priority patent/US4683195A/en
Priority claimed from DK13296A external-priority patent/DK175469B1/en
Application filed by Hoffmann La Roche filed Critical Hoffmann La Roche
Publication of DK200401109A publication Critical patent/DK200401109A/en
Application granted granted Critical
Publication of DK175778B1 publication Critical patent/DK175778B1/en

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Description

PATENTKRAV 1. Eksponentielt multiplicerings- og påvisningskit til multiplicering og påvisning af en specifik template-nukleinsyresekvens eller specifikke template-nukleinsyresekvenser, der er indeholdt i en enkelt- eller dobbeltstrenget nukleinsyre eller i en blanding af sådanne nukleinsyrer i en prøve, hvilket kit i samlet form omfatter: (a) i det mindste en første og en anden oligonukleotidprimer, der er indbyrdes forskellige, hvori (aa) den ene af primerne er i det væsentlige komplementær til den enkeltstrengede nukleinsyre eller til en streng i den dobbeltstrengede nukleinsyre, (ab) den anden af primerne er i det væsentlige komplementær til et komplement af den enkeltstrengede nukleinsyre eller til den anden streng i den dobbeltstrengede nukleinsyre, og hvori (ac) primerne afgrænser enderne af den specifikke nukleinsyresekvens, der skal multipliceres og detekteres; (b) et polymeriseringsmiddel; og (c) midler til detektering af den multiplicerede specifikke nukleinsyresekvens. 2. Kit ifølge krav 1, hvori den specifikke template-nukleinsyresekvens er indeholdt i en større sekvens. 3. Kit ifølge krav 1 eller 2, hvori nukleinsyren er DNA eller RNA, herunder mRNA, hvilket DNA eller RNA kan være enkelt- eller dobbeltstrenget eller er en DNA-RNA-hybrid. 4. Kit ifølge et hvilket som helst af kravene 1 til 3, hvori nukleinsyren er genomisk DNA. 5. Kit ifølge et hvilket som helst af kravene 1 til 4, hvori primerne indeholder ca. 15 til 25 nukleotider. 6. Kit ifølge et hvilket som helst af kravene 1 til 5, hvori mindst den ene af primerne har en nukleotidsekvens bundet til sin S'-ende, hvilken sekvens er ikke-kom-plementær til nukleinsyren. 7. Kit ifølge krav 6, hvori nukleotidsekvensen, der er bundet til 5'-enden af primeren, er en promotorsekvens. 8. Kit ifølge krav 7, hvori promotoren er T7-promotoren. 9. Kit ifølge et hvilket som helst af kravene 1-5, hvori der spredt i det mindste i den ene af primerne er anbragt baser eller en nukleotidsekvens, hvilke baser eller hvilken nukleotidsekvens er ikke-komplementære til den specifikke nukleinsyresekvens, der skal multipliceres og detekteres. 10. Kit ifølge et hvilket som helst af kravene 1 til 5, hvori den ene primer indeholder et restriktionssted. 11. Kit ifølge et hvilket som helst af kravene 1 til 10, hvori polymeriseringsmidlet er et enzym valgt blandt E. coli polymerase 1, Klenow-fragmentet fra E. coli polymerase J, T4-DNA-polymerase, andre DNA-polymeraser, revers transkriptase og varmestabile enzymer. 12. Kit ifølge et hvilket som helst af kravene 1 til 11, hvori midlet til detektering af den multiplicerede specifikke template-nukleinsyresekvens eller de multiplicerede specifikke template-nukleinsyresekvenser omfatter en mærket oligonukleotidprobe. 13. Anvendelse af et kit ifølge et hvilket som helst af kravene 1 til 12 til multiplikation, detektering og/eller karakterisering afen specifik template-nukleinsyresekvens eller specifikke template-nukieinsyresekvenser indeholdt i en enkelt- eller dobbeltstrenget nukleinsyre eller i en blanding af sådanne nukleinsyrer. 14. Anvendelse ifølge krav 13, hvori den specifikke template-nukleinsyresekvens eller de specifikke template-nukleinsyresekvenser er associeret med infektiøse sygdomme såsom dem, der er forårsaget af bakterier, vira og protozoparasitter, genetiske lidelser såsom dem, der er forårsaget af specifikke deletioner og/eller mutationer i genomisk DNA, eller cellulære lidelser såsom cancer.PATENT REQUIREMENT 1. Exponential multiplication and detection kit for multiplying and detecting a specific template nucleic acid sequence or specific template nucleic acid sequences contained in a single or double stranded nucleic acid or in a mixture of such nucleic acids in a sample, which kit in aggregate form comprises: (a) at least a first and second oligonucleotide primer that are mutually different, wherein (aa) one of the primers is substantially complementary to the single-stranded nucleic acid or to a strand of the double-stranded nucleic acid, (ab) the second of the primers is substantially complementary to a complement of the single-stranded nucleic acid or to the second strand of the double-stranded nucleic acid, wherein the (ac) primers define the ends of the specific nucleic acid sequence to be multiplied and detected; (b) a polymerizing agent; and (c) means for detecting the multiplied specific nucleic acid sequence. The kit of claim 1, wherein the specific template nucleic acid sequence is contained in a larger sequence. The kit of claim 1 or 2, wherein the nucleic acid is DNA or RNA, including mRNA, which DNA or RNA may be single or double stranded or is a DNA-RNA hybrid. The kit of any one of claims 1 to 3, wherein the nucleic acid is genomic DNA. The kit of any one of claims 1 to 4, wherein the primers contain approx. 15 to 25 nucleotides. The kit of any one of claims 1 to 5, wherein at least one of the primers has a nucleotide sequence bound to its S 'end, which sequence is non-complementary to the nucleic acid. The kit of claim 6, wherein the nucleotide sequence bound to the 5 'end of the primer is a promoter sequence. The kit of claim 7, wherein the promoter is the T7 promoter. A kit according to any one of claims 1-5, wherein at least one of the primers dispersed comprises bases or a nucleotide sequence, which bases or nucleotide sequence are non-complementary to the specific nucleic acid sequence to be multiplied and detected. The kit of any one of claims 1 to 5, wherein said one primer contains a restriction site. The kit of any one of claims 1 to 10, wherein the polymerizing agent is an enzyme selected from E. coli polymerase 1, the Klenow fragment of E. coli polymerase J, T4 DNA polymerase, other DNA polymerases, reverse transcriptase and heat-stable enzymes. The kit of any one of claims 1 to 11, wherein the means for detecting the multiplied specific template nucleic acid sequence or the multiplied specific template nucleic acid sequences comprises a labeled oligonucleotide probe. Use of a kit according to any one of claims 1 to 12 for multiplication, detection and / or characterization of a specific template nucleic acid sequence or specific template nucleic acid sequences contained in a single or double stranded nucleic acid or in a mixture of such nucleic acids. Use according to claim 13, wherein the specific template nucleic acid sequence or specific template nucleic acid sequences are associated with infectious diseases such as those caused by bacteria, viruses and protozoparasites, genetic disorders such as those caused by specific deletions and / or mutations in genomic DNA, or cellular disorders such as cancer.

DK200401109A 1985-03-28 2004-07-15 Kit for amplification and detection of specific nucleic acid sequences - used to characterise or detect sequences associated with infectious diseases, genetic disorders and cellular disorders DK175778B1 (en)

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
US71697585A 1985-03-28 1985-03-28
US71697585 1985-03-28
US06/791,308 US4683202A (en) 1985-03-28 1985-10-25 Process for amplifying nucleic acid sequences
US79130885 1985-10-25
US06/828,144 US4683195A (en) 1986-01-30 1986-02-07 Process for amplifying, detecting, and/or-cloning nucleic acid sequences
US82814486 1986-02-07
DK13296 1996-02-09
DK13296A DK175469B1 (en) 1985-03-28 1996-02-09 Method for Multiplication and Detection of Nucleic Acid Sequences

Publications (2)

Publication Number Publication Date
DK200401109A true DK200401109A (en) 2004-07-15
DK175778B1 DK175778B1 (en) 2005-02-14

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DK200401109A DK175778B1 (en) 1985-03-28 2004-07-15 Kit for amplification and detection of specific nucleic acid sequences - used to characterise or detect sequences associated with infectious diseases, genetic disorders and cellular disorders

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DK (1) DK175778B1 (en)

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DK175778B1 (en) 2005-02-14

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