Real-time detection of BRCA1 gene mutations using a monolithic silicon optocoupler array.
The monolithic silicon optocoupler presented here offers a platform for a new generation of fully integrated devices fabricated through the mature and inexpensive silicon technology. Using the developed optocoupler, real-time detection of the most common mutations in BRCA1 gene related to predisposition to hereditary breast/ovarian cancer was accomplished. For this purpose, oligonucleotides corresponding to wild- and mutant-type sequences were immobilized onto different optocouplers and the hybridization with fluorescently labeled complementary or non-complementary sequences was monitored in real time. Hybridization of fluorescently labeled oligonucleotides to the immobilized ones modulated the coupling efficiency between the light emitting diode and the detector in a concentration-dependent manner. Using as label the AlexaFluor 647 dye (whose absorption maximum fits the emission maximum of the light source) a detection limit of 0.9 nM (9 fmol) was achieved. Real-time signal monitoring, especially during dehybridization, improved considerably the discrimination between wild-type and mutant sequences due to the ability to calculate dissociation kinetics upon washing independently for each one mutation. The bioanalytical capabilities of the transducer along with the fact that dense transducer arrays can be fabricated on a single chip open new frontiers in the manufacturing of microsystems appropriate for point-of-care analysis.
Mavrogiannopoulou
Elissavet
E
Immunoassay/Immunosensors Lab., I./R.-R.P., N.C.S.R. "Demokritos", GR-15310 Aghia Paraskevi, Greece.
Petrou
Panagiota S
PS
Kakabakos
Sotirios E
SE
Misiakos
Konstantinos
K
D019313:BRCA1 Protein / Q000235:genetics*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D003199:Computer Systems; D004252:DNA Mutational Analysis / Q000379:methods*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D019398:Genes, BRCA1; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation*; D055096:Optical Devices; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012825:Silicon / Q000737:chemistry*
D019313:BRCA1 Protein; D012825:Silicon