ES2173782B1 - QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN COMPETITIVE LIGASA CHAIN. - Google Patents
QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN COMPETITIVE LIGASA CHAIN.Info
- Publication number
- ES2173782B1 ES2173782B1 ES009902758A ES9902758A ES2173782B1 ES 2173782 B1 ES2173782 B1 ES 2173782B1 ES 009902758 A ES009902758 A ES 009902758A ES 9902758 A ES9902758 A ES 9902758A ES 2173782 B1 ES2173782 B1 ES 2173782B1
- Authority
- ES
- Spain
- Prior art keywords
- analyte
- amplification
- ligands
- standard
- competitive
- 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.)
- Expired - Fee Related
Links
Abstract
Procedimiento de cuantificación de ácidos nucleicos mediante amplificación por reacción en cadena de ligasa competitiva, basado en la co-amplificación de un segmento do ácido nucleico analito y un segmento de ácido nucleico patrón intrínsecamente ajeno a la muestra que contiene el analito, siendo dicha co- amplificación realizada mediante una reacción en cadena de ligasa (LCR) en la que intervienen seis oligonuceótidos ligando, A1, A2, P1, P2, C1 y C2, siendo el analito amplificado por el cuarteto formado por los ligandos A1, A2, C1 y C2, y el patrón amplificado por el cuarteto P1, P2, C1 y C2, siendo A1 y A2 ligandos específicos y exclusivos del analito, P1 y P2 ligandos específicos y exclusivos del patrón, y C1 y C2 ligandos específicos de forma común para el analito y el patrón, de forma que durante el proceso de amplificación se establece una competición por la unión de los ligandos C1 y C2 entre el analito y el patrón, o sus respectivos productos de amplificación. La cuantificación delanalito se consigue añadiendo una cantidad conocida del patrón a una alícuota de la muestra que se sospecha que contiene dicho analito, realizando una LCR con los ligandos A1, A2, P1, P2, C1 y C2, cuantificando los productos de amplificación del analito y del patrón, y deduciendo la cantidad de analito presente en la muestra inicial a partir de las cantidades relativas de dichos productos de amplificación.Nucleic acid quantification process by amplification by competitive ligase chain reaction, based on the co-amplification of an analyte nucleic acid segment and an intrinsically foreign standard nucleic acid segment to the sample containing the analyte, said co- amplification performed by a ligase chain reaction (LCR) in which six ligand oligonucleotides, A1, A2, P1, P2, C1 and C2 are involved, the analyte being amplified by the quartet formed by ligands A1, A2, C1 and C2 , and the pattern amplified by quartet P1, P2, C1 and C2, with A1 and A2 being specific and exclusive ligands of the analyte, P1 and P2 specific and exclusive ligands of the pattern, and C1 and C2 specific ligands in common form for the analyte and the standard, so that during the amplification process a competition is established for the union of ligands C1 and C2 between the analyte and the standard, or their respective amplification products ation The analyte quantification is achieved by adding a known amount of the standard to an aliquot of the sample suspected of containing said analyte, performing a CSF with ligands A1, A2, P1, P2, C1 and C2, quantifying the analyte amplification products and of the standard, and deducting the amount of analyte present in the initial sample from the relative amounts of said amplification products.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES009902758A ES2173782B1 (en) | 1999-12-17 | 1999-12-17 | QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN COMPETITIVE LIGASA CHAIN. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES009902758A ES2173782B1 (en) | 1999-12-17 | 1999-12-17 | QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN COMPETITIVE LIGASA CHAIN. |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| ES2173782A1 ES2173782A1 (en) | 2002-10-16 |
| ES2173782B1 true ES2173782B1 (en) | 2003-12-16 |
Family
ID=8310939
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES009902758A Expired - Fee Related ES2173782B1 (en) | 1999-12-17 | 1999-12-17 | QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN COMPETITIVE LIGASA CHAIN. |
Country Status (1)
| Country | Link |
|---|---|
| ES (1) | ES2173782B1 (en) |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB9116042D0 (en) * | 1991-07-24 | 1991-09-11 | Royal Free Hosp School Med | Diagnostic method |
| WO1995006137A1 (en) * | 1993-08-27 | 1995-03-02 | Australian Red Cross Society | Detection of genes |
| NO301082B1 (en) * | 1995-07-20 | 1997-09-08 | Statens Inst For Folkehelse | Method of monitoring the result in Gap-LCR, as well as nucleic acid sequence for use in the method |
-
1999
- 1999-12-17 ES ES009902758A patent/ES2173782B1/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| ES2173782A1 (en) | 2002-10-16 |
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