DNA analysis and diagnostics on oligonucleotide microchips.
We present a further development in the technology of sequencing by hybridization to oligonucleotide microchips (SHOM) and its application to diagnostics for genetic diseases. A robot has been constructed to manufacture sequencing "microchips." The microchip is an array of oligonucleotides immobilized into gel elements fixed on a glass plate. Hybridization of the microchip with fluorescently labeled DNA was monitored in real time simultaneously for all microchip elements with a two-wavelength fluorescent microscope equipped with a charge-coupled device camera. SHOM has been used to detect beta-thalassemia mutations in patients by hybridizing PCR-amplified DNA with the microchips. A contiguous stacking hybridization technique has been applied for the detection of mutations; it can simplify medical diagnostics and enhance its reliability. The use of multicolor monitoring of contiguous stacking hybridization is suggested for large-scale diagnostics and gene polymorphism studies. Other applications of the SHOM technology are discussed.
Yershov
G
G
Joint Human Genome Program: Engelhardt Institute of Molecular Biology, Moscow, Russia.
Barsky
V
V
Belgovskiy
A
A
Kirillov
E
E
Kreindlin
E
E
Ivanov
I
I
Parinov
S
S
Guschin
D
D
Drobishev
A
A
Dubiley
S
S
Mirzabekov
A
A
D001483:Base Sequence; D001709:Biotechnology; D004247:DNA / Q000235:genetics; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D005069:Evaluation Studies as Topic; D030342:Genetic Diseases, Inborn / Q000175:diagnosis* / Q000235:genetics; D005821:Genetic Techniques / Q000295:instrumentation; D005914:Globins / Q000235:genetics; D006801:Humans; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D009841:Oligonucleotides / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012371:Robotics / Q000295:instrumentation; D017086:beta-Thalassemia / Q000175:diagnosis / Q000235:genetics
D017931:DNA Primers; D009841:Oligonucleotides; D005914:Globins; D004247:DNA