Affinity sensing for transgenes detection in antidoping control.
Sports authorities fear that a new form of doping called gene doping, based on the misuse of gene therapy, represents an emerging important problem and so far no methods are available for detecting it. The World Anti-Doping Agency (WADA) has included since 2003 for the first time gene doping methods in the "Prohibited List of Substances and Methods", thus detection of this new form of doping is challenging for analytical chemists. In this work, we apply affinity-based biosensors (ABBs), in particular DNA piezoelectric sensing, for detection of target DNA sequences selected as transgenosis markers. In this work, two sequences widely used in transgenosis experiments have been identified as markers: the enhanced green fluorescence protein (EGFP) gene and the promoter of Cytomegalovirus (CMV). The biosensors are characterized in their analytical performances using synthetic oligonucleotides and amplified DNA obtained from purified plasmid used as a template. Finally they have been applied to transgenic human cell cultures (human embryonic kidney HEK-EGFP), transformed with the same plasmid and carrying the target markers. This represents the closest human real matrix available for our transgenes.
Scarano
Simona
S
Dipartimento di Chimica, Università degli Studi di Firenze, 50019 Sesto Fiorentino, Italy.
Spiriti
Maria M
MM
Tigli
Genny
G
Bogani
Patrizia
P
Buiatti
Marcello
M
Mascini
Marco
M
Minunni
Maria
M
D001483:Base Sequence; D015374:Biosensing Techniques / Q000379:methods*; D002138:Calibration; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics; D004247:DNA / Q000235:genetics; D015342:DNA Probes / Q000138:chemical synthesis / Q000737:chemistry / Q000235:genetics; D004300:Doping in Sports; D004560:Electricity; D005819:Genetic Markers / Q000235:genetics; D015316:Genetic Therapy; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics; D019076:Transgenes
D015342:DNA Probes; D005819:Genetic Markers; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D004247:DNA