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Comparison of DNA amplification methods for simultaneous detection of Salmonella spp. and Cronobacter sakazakii in milk

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Comparison of DNA amplification methods for simultaneous detection of Salmonella spp. and Cronobacter sakazakii in milk

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dc.contributor.advisor Tortajada Genaro, Luis Antonio es_ES
dc.contributor.advisor Puchades Pla, Rosa es_ES
dc.contributor.advisor Maquieira Catala, Ángel es_ES
dc.contributor.author Santiago Felipe, Sara es_ES
dc.date.accessioned 2016-02-04T19:23:24Z
dc.date.available 2016-02-04T19:23:24Z
dc.date.created 2012-07
dc.date.issued 2016-02-04
dc.identifier.uri http://hdl.handle.net/10251/60645
dc.description.abstract The control and detection of foodborne pathogens is a major problem in public health. Therefore, the development of rapid and economical diagnostic method along the food chain has become a priority. Among  the  available  methodologies,  those  based  on  the  detection  of  nucleic  acids provide certain advantages such as high sensitivity speed, selectivity, and the ability  to  simultaneously  detect  multiple  microorganisms.  However,  the  amount  of  pathogen DNA present in a food is minimal, so these methods must incorporate an amplification  stage  that  achieves  the  proper  amount  for  their  detection.  The  polymerase chain reaction (PCR) is the common technique, although this method has the requirement of thermocycling. In this master project two enzymatic reactions that enable the amplification of DNA  under  isothermal  conditions  have  been  studied.  The  work  focuses  on  the  comparison of conventional PCR with two isothermal amplification methods ‐recombinase polymerase (RPA) and multiple displacement amplification (MDA)‐. These methods, applied to the detection of Salmonella spp. and Cronobacter sakazakii, have shown excellent amplification yields of the target sequences. In addition, the inhibitory effect of some matrix components on the enzymes has been studied. The simultaneous detection of two pathogens has been made in milk samples, analyzing  the  amplified products  by  hybridization  assays  with  oligonucleotide  probes  immobilized on the surface of a DVD in microarray format. The results have been satisfactory in terms of reproducibility and sensitivity. It  has  been  shown  that  the  use  of  other  alternative  enzymes  and  low  cost  heating equipments allow DNA amplification at constant temperature. Moreover, the combination  with  compact  disc  technology  increases  the  advantages,  demonstrating  the validity of the proposed biosensor in routine applications in the area of food safety. es_ES
dc.format.extent 26 es_ES
dc.language Inglés es_ES
dc.publisher Universitat Politècnica de València es_ES
dc.rights Reserva de todos los derechos es_ES
dc.subject DNA amplification methods es_ES
dc.subject Multiple displacement amplification (MDA) es_ES
dc.subject Recombinase Polymerase Amplification (RPA) es_ES
dc.subject Cronobacter Sakazakii es_ES
dc.subject Salmonella spp. es_ES
dc.subject.classification QUIMICA ANALITICA es_ES
dc.subject.other Máster Universitario en Sensores para Aplicaciones Industriales-Màster Universitari en Sensors per a Aplicacions Industrials es_ES
dc.title Comparison of DNA amplification methods for simultaneous detection of Salmonella spp. and Cronobacter sakazakii in milk es_ES
dc.type Tesis de máster es_ES
dc.rights.accessRights Cerrado es_ES
dc.contributor.affiliation Universitat Politècnica de València. Escuela Técnica Superior de Ingeniería del Diseño - Escola Tècnica Superior d'Enginyeria del Disseny es_ES
dc.description.bibliographicCitation Santiago Felipe, S. (2012). Comparison of DNA amplification methods for simultaneous detection of Salmonella spp. and Cronobacter sakazakii in milk. http://hdl.handle.net/10251/60645 es_ES
dc.description.accrualMethod Archivo delegado es_ES


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