An electrochemical detection scheme for identification of single nucleotide polymorphisms using hairpin-forming probes.
Single nucleotide polymorphisms are implicated as having a significant role in regulating growth, development and, thereby, human health and disease. We have developed a method for identifying single nucleotide genetic alterations by combining hairpin-forming DNA probes and electrochemical detection of sandwich DNA hybridization. Incorporation of hairpin-forming competitor probes and the catalyzed reporter deposition amplification system further improves assay specificity by 7-fold and sensitivity by 100-fold. We have demonstrated that the system successfully identified the factor V Leiden mutations from human blood specimens.
Huang
Tony Jun
TJ
Mechanical and Aerospace Engineering Department, University of California-Los Angeles, Los Angeles, CA 90095-1597, USA.
Liu
Minghsun
M
Knight
Linda D
LD
Grody
Wayne W
WW
Miller
Jeff F
JF
Ho
Chih-Ming
CM
D015342:DNA Probes / Q000737:chemistry*; D004563:Electrochemistry / Q000379:methods*; D005165:Factor V / Q000235:genetics; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods; D009690:Nucleic Acid Conformation; D015345:Oligonucleotide Probes / Q000737:chemistry; D009841:Oligonucleotides / Q000032:analysis; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA / Q000379:methods*
D015342:DNA Probes; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; C095381:factor V Leiden; D005165:Factor V