RT Journal Article
T1 Disposable Amperometric Polymerase Chain Reaction-FreeBiosensor for Direct Detection of Adulteration with Horsemeat in Raw Lysates Targeting Mitochondrial DNA
A1 Ruiz-Valdepeñas Montiel, Victor
A1 Gutiérrez, María L.
A1 Povedano, Eloy
A1 Torrente Rodríguez, Rebeca Magnolia
A1 Vargas, Eva
A1 Reviejo García, Ángel Julio
A1 Linacero de la Fuente, Rosario
A1 Gallego, Francisco J.
A1 Campuzano Ruiz, Susana
A1 Pingarrón Carrazón, José Manuel
AB A novel electrochemical disposable nucleic acid biosensor for simple, rapid, and specific detection of adulterations with horsemeat is reported in this work. The biosensing platform involves immobilization of a 40-mer RNA probe specific for a characteristic fragment of the mitochondrial DNA D-loop region of horse onto the surface of magnetic microcarriers. In addition, signal amplification was accomplished by using a commercial antibody specific to RNA/ DNA duplexes and a bacterial protein conjugated with a horseradish peroxidase homopolymer (ProtA-HRP40). Amperometric detection at −0.20 V vs Ag pseudoreference electrode was carried out at disposable screen-printed carbon electrodes. The methodology achieved a limit of detection (LOD) of 0.12 pM (3.0 attomoles) for the synthetic target and showed ability to discriminate between raw beef and horsemeat using just 50 ng of total extracted mitochondrial DNA (∼16 660 bp in length) without previous fragmentation. The biosensor also allowed discrimination between 100% raw beef and beef meat samples spiked with only 0.5% (w/w) horse meat (levels established by the European Commission) using raw mitochondrial lysates without DNA extraction or polymerase chain reaction (PCR) amplification in just 75 min. These interesting features made the developed methodology an extremely interesting tool for beef meat screening, and it can be easily adapted to the determination of other meat adulterations by selection of the appropriate specific fragments of the mitochondrial DNA region and capture probes.
PB ACS
SN (Print Edition) 0003-2700 (Web Edition) 1520-6882
YR 2017
FD 2017
LK https://hdl.handle.net/20.500.14352/18078
UL https://hdl.handle.net/20.500.14352/18078
LA eng
NO Ministerio de Economía y Competitividad (MINECO)
NO Comunidad de Madrid
DS Docta Complutense
RD 23 abr 2025