Genosensor based on a platinum(II) complex as electrocatalytic label.
Voltammetric genosensors on streptavidin-modified screen-printed carbon electrodes (SPCEs) for the detection of virulence nucleic acid determinants of pneumolysin (ply) and autolysin (lytA) genes, exclusively present on the genome of the human pathogen Streptococcus pneumoniae, were described. The oligonucleotide probes were immobilized on electrochemically pretreated SPCEs through the streptavidin/biotin reaction. After that, the hybridization reaction was carried out with labeled complementary targets on the electrode surface. The ply and lytA targets were labeled using the universal linkage system, which consists of the use of a platinum(II) complex that acts as coupling agent between targets and a, usually fluorescent, molecule label. In this case, the platinum(II) complex acts as a label itself because the analytical signal is achieved by measuring chronoamperometrically the current generated by the hydrogen evolution catalyzed by platinum. In nonstringent experimental conditions, these genosensors can detect 24.5 fmol of 20-mer oligonucleotide target and discriminate between a complementary oligo and an oligo with a three-base mismatch. In presence of 25% formamide in the hybridization buffer, a single-base mismatch on the oligonucleotide target can be detected.
Hernández-Santos
David
D
Departamento de Química Física y Analítica, Facultad de Química, Universidad de Oviedo. 33006 Oviedo, Asturias, Spain.
González-García
María Begoña
MB
Costa-García
Agustín
A
D001426:Bacterial Proteins / Q000032:analysis / Q000235:genetics; D002384:Catalysis; D004563:Electrochemistry / Q000379:methods*; D004566:Electrodes; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000032:analysis / Q000235:genetics*; D009693:Nucleic Acid Hybridization / Q000379:methods; D017671:Platinum Compounds / Q000737:chemistry*; D019809:Streptavidin / Q000737:chemistry; D013296:Streptococcus pneumoniae / Q000235:genetics*; D013301:Streptolysins / Q000032:analysis / Q000235:genetics*
D001426:Bacterial Proteins; D017671:Platinum Compounds; D013301:Streptolysins; C032290:plY protein, Streptococcus pneumoniae; D019809:Streptavidin; D009238:N-Acetylmuramoyl-L-alanine Amidase