ES2170620B1 - QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN HEMI-COMPETITIVE POLYMERASE CHAIN. - Google Patents
QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN HEMI-COMPETITIVE POLYMERASE CHAIN.Info
- Publication number
- ES2170620B1 ES2170620B1 ES009902600A ES9902600A ES2170620B1 ES 2170620 B1 ES2170620 B1 ES 2170620B1 ES 009902600 A ES009902600 A ES 009902600A ES 9902600 A ES9902600 A ES 9902600A ES 2170620 B1 ES2170620 B1 ES 2170620B1
- Authority
- ES
- Spain
- Prior art keywords
- analyte
- amplification
- standard
- polymerase chain
- primer
- 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 polimerasa hemi- competitiva, basado en la co-amplificación de un segmento de ácido nucleico analito y un segmento de ácido nucleico patrón intrínsecamente ajeno a la muestra que contiene el analito, siendo dicha coamplificación realizada mediante una reacción en cadena de polimerasa (PCR) en la que intervienen tres oligonuceótidos cebadores, A, B y C, siendo el analito amplificado por la pareja formada por los cebadores A y C, y el patrón amplificado por la pareja B y C, siendo A un cebador específico y exclusivo del analito, B un cebador específico y exclusivo del patrón, y C un cebador especifico de forma común para el analito y el patrón, de forma que durante el proceso de amplificación se establece una competición por la unión del cebador C entre el analito y el patrón, o sus respectivos productos de amplificación. La cuantificación del analito se consigue añadiendo una cantidad conocida del patrón a una alícuota de la muestra que se sospecha que contiene dicho analito, realizando una PCR con los cebadores A, B y C, 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 hemi-competitive polymerase 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 coamplification performed by a polymerase chain reaction (PCR) in which three oligonucleotide primers, A, B and C, are involved, the analyte being amplified by the pair formed by primers A and C, and the pattern amplified by the pair B and C, being A a specific and exclusive primer of the analyte, B a specific and exclusive primer of the standard, and C a specific primer commonly for the analyte and the standard, so that during the amplification process a competition for the binding of primer C between the analyte and the standard, or their respective amplification products. Quantification of the analyte is achieved by adding a known amount of the standard to an aliquot of the sample suspected of containing said analyte, performing a PCR with primers A, B and C, quantifying the amplification products of the analyte and 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 |
|---|---|---|---|
| ES009902600A ES2170620B1 (en) | 1999-11-25 | 1999-11-25 | QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN HEMI-COMPETITIVE POLYMERASE CHAIN. |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| ES009902600A ES2170620B1 (en) | 1999-11-25 | 1999-11-25 | QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN HEMI-COMPETITIVE POLYMERASE CHAIN. |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| ES2170620A1 ES2170620A1 (en) | 2002-08-01 |
| ES2170620B1 true ES2170620B1 (en) | 2003-12-16 |
Family
ID=8310722
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| ES009902600A Expired - Fee Related ES2170620B1 (en) | 1999-11-25 | 1999-11-25 | QUANTIFICATION PROCEDURE OF NUCLEIC ACIDS THROUGH AMPLIFICATION BY REACTION IN HEMI-COMPETITIVE POLYMERASE CHAIN. |
Country Status (1)
| Country | Link |
|---|---|
| ES (1) | ES2170620B1 (en) |
Family Cites Families (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| FR2668162B1 (en) * | 1990-10-17 | 1993-01-22 | Eurobio Lab | DNA FRAGMENTS, METHOD AND DIAGNOSTIC KIT FOR DETECTION OF DELTA-F-508 MUTATION CARRIERS RESPONSIBLE FOR MUCOVISCIDOSIS. |
| WO1993018177A1 (en) * | 1992-03-13 | 1993-09-16 | The Children's Hospital Of Philadelphia | Diagnosis of cystic fibrosis using allele specific multiplex polymerase chain reactions |
| ATE198358T1 (en) * | 1992-04-27 | 2001-01-15 | Dartmouth College | DETECTION OF GENE SEQUENCES IN BIOLOGICAL LIQUIDS |
| WO1995006137A1 (en) * | 1993-08-27 | 1995-03-02 | Australian Red Cross Society | Detection of genes |
| US5667974A (en) * | 1995-06-07 | 1997-09-16 | Abbott Laboratories | Method for detecting nucleic acid sequences using competitive amplification |
-
1999
- 1999-11-25 ES ES009902600A patent/ES2170620B1/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| ES2170620A1 (en) | 2002-08-01 |
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Legal Events
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