ES2543172T3 - Sonda de ácido nucleico peptídico, estuche y método para la detección y/o cuantificación de Salmonella spp. y sus aplicaciones - Google Patents

Sonda de ácido nucleico peptídico, estuche y método para la detección y/o cuantificación de Salmonella spp. y sus aplicaciones Download PDF

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ES2543172T3
ES2543172T3 ES11719888.7T ES11719888T ES2543172T3 ES 2543172 T3 ES2543172 T3 ES 2543172T3 ES 11719888 T ES11719888 T ES 11719888T ES 2543172 T3 ES2543172 T3 ES 2543172T3
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detection
quantification
kit
applications
nucleic acid
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Nuno Filipe Ribeiro Pinto De Oliveira Azevedo
Maria João LOPES DA COSTA VIEIRA
Carina Manuela Fernandes Almeida
Charles William Keevil
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Universidade do Minho
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Universidade do Minho
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    • 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
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    • 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
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • C—CHEMISTRY; METALLURGY
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    • 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
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    • 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
    • C12Q1/6811—Selection methods for production or design of target specific oligonucleotides or binding molecules
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    • 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
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    • 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
    • C12Q1/6851—Quantitative amplification
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    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/107—Modifications characterised by incorporating a peptide nucleic acid
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    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/42—Salmonella
    • 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
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

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  • Chemical & Material Sciences (AREA)
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  • Genetics & Genomics (AREA)
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  • Toxicology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

Sonda de PNA para la detección y/o cuantificación de Salmonella, caracterizada por que comprende SEQ ID No. 1 - 5'-AGG AGC TTC GCT TGC-3'.

Description

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Tabla 1 – Especificidades y sensibilidades teóricas de las sondas existentes para la detección de Salmonella spp.
Sonda Sal3 SEQ No.8 Salm63 SEQ No.7 Sal23S10 SEQ No.6 SalPNA1873 SEQ No. 1
Tipo ADN ADN PNA PNA
Secuencia (5’3’) AATCACTTCACCTAC GTG TCGACTGACTTCAGCT CC TAAGCCGGGATG GC AGGAGCTTCGCTT GC
N° de cepas de Salmonella detectadas 96* 73* 95* 101*
N° of non-Salmonella detectadas 0# 3# 206# 0#
Especificidad (%) 100 99,97 98,13 100
Sensibilidad (%) 95,05 72,23 94,06 100
Referencia Nordentoft y cols., 1997 Kutter y cols., 2006 O’ Keefe y cols., 2001 Este trabajo
* Cepas de Salmonella detectadas en un total de 101 secuencias de Salmonella presentes en la base de datos. # Cepas que no son de Salmonella detectadas en un total de 11.023 secuencias que no son de Salmonella depositadas en la base de datos.
La sonda de PNA de esta invención comprende preferiblemente 15 nucleótidos y es idéntica a la secuencia SEQ ID 5 No. 1 -5’-AGG AGC TTC GCT TGC-3’.
AlteARNtivamente, esta invención también contempla variaciones de las secuencias de nucleótidos de las sondas.
Fracción detectable de la sonda de PNA:
No limitada a los siguientes ejemplos, la fracción detectable de sonda de PNA puede incluir diversos tipos de moléculas tales como conjugados de dextrano, cromóforos, fluoróforos, radioisótopos, enzimas, haptenos, un 10 compuesto quimioluminiscente, entre otras.
Como un ejemplo, entre la clase de fluoróforos, los que son preferibles para usar son (pero no se limitan a): fluoróforos de la serie Alexa, la serie Alexa Fluor, cianinas, 5-(y 6)-carboxi-2’,7’-diclorofluoresceína, 5-ROX (5carboxi-X-rodamina, sal de trietilamonio).
Método:
15 La presente invención presenta un método para determinar la presencia de Salmonella usando una secuencia de nucleótidos con al menos 86% de homología con la región de 15 nucleótidos descrita en la presente -SEQ ID No. 1.
El método puede incluir el contacto de una muestra con una sonda de PNA descrita en la presente con la secuencia diana de la bacteria en condiciones de hibridación apropiadas o condiciones de hibridación in situ apropiadas (según se muestra en el EJEMPLO 1).
20 El método se puede dividir en: preparación de muestra (que incluye la etapa de enriquecimiento, cuando fuera necesaria), fijación, hibridación, lavado y visualización de los resultados (véase el EJEMPLO 1).
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Claims (1)

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    imagen2
ES11719888.7T 2010-03-29 2011-03-29 Sonda de ácido nucleico peptídico, estuche y método para la detección y/o cuantificación de Salmonella spp. y sus aplicaciones Active ES2543172T3 (es)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
PT105029A PT105029B (pt) 2010-03-29 2010-03-29 Sonda de ácido péptido nucleico, estojo e método para detectar e/ou quantificar salmonella e respectivas aplicações
PT10005029 2010-03-29
PCT/IB2011/051337 WO2011121544A1 (en) 2010-03-29 2011-03-29 Peptide nucleicacid probe, kit and method for detection and/or quantification of salmonella spp. and applications thereof

Publications (1)

Publication Number Publication Date
ES2543172T3 true ES2543172T3 (es) 2015-08-17

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ID=44168263

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ES11719888.7T Active ES2543172T3 (es) 2010-03-29 2011-03-29 Sonda de ácido nucleico peptídico, estuche y método para la detección y/o cuantificación de Salmonella spp. y sus aplicaciones

Country Status (11)

Country Link
US (1) US20130102000A1 (es)
EP (1) EP2553120B1 (es)
JP (1) JP2013523118A (es)
KR (1) KR20130062276A (es)
CN (1) CN102939390B (es)
BR (1) BR112012024873A2 (es)
CA (1) CA2794466A1 (es)
ES (1) ES2543172T3 (es)
PT (1) PT105029B (es)
RU (1) RU2567011C2 (es)
WO (1) WO2011121544A1 (es)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102586466B (zh) * 2012-03-23 2014-02-26 浙江省检验检疫科学技术研究院 肽核酸原位荧光杂交技术检测沙门氏菌的方法及pna探针
CN104342493B (zh) * 2014-10-13 2016-12-07 浙江省检验检疫科学技术研究院 单核细胞增多性李斯特菌的肽核酸分子信标标记和荧光扫描检测方法

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6664045B1 (en) * 1998-06-18 2003-12-16 Boston Probes, Inc. PNA probes, probe sets, methods and kits pertaining to the detection of microorganisms
DE60138916D1 (de) 2000-09-26 2009-07-16 Boston Probes Inc Sonden, sondensätzen,verfahren und kits zur nachweis, identifizierung und/oder die zählung von bakterien
AU2002255478A1 (en) * 2001-01-10 2002-09-12 Pe Corporation (Ny) Kits, such as nucleic acid arrays, comprising a majority of human exons or transcripts, for detecting expression and other uses thereof

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Publication number Publication date
US20130102000A1 (en) 2013-04-25
WO2011121544A1 (en) 2011-10-06
KR20130062276A (ko) 2013-06-12
JP2013523118A (ja) 2013-06-17
WO2011121544A4 (en) 2011-12-01
PT105029A (pt) 2011-09-29
PT105029B (pt) 2012-09-11
RU2567011C2 (ru) 2015-10-27
CN102939390B (zh) 2016-05-18
BR112012024873A2 (pt) 2016-12-06
EP2553120A1 (en) 2013-02-06
EP2553120B1 (en) 2015-04-22
CN102939390A (zh) 2013-02-20
CA2794466A1 (en) 2011-10-06
RU2012141287A (ru) 2014-05-10

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