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Nucleic Acids Research, 2002, Vol. 30, No. 12 e55
© 2002 Oxford University Press

An electrochemical detection scheme for identification of single nucleotide polymorphisms using hairpin-forming probes

Tony Jun Huang, Minghsun Liu1, Linda D. Knight2, Wayne W. Grody2,3, Jeff F. Miller1 and Chih-Ming Ho*

Mechanical and Aerospace Engineering Department, 1Department of Microbiology, Immunology and Molecular Genetics, 2Department of Pathology and Laboratory Medicine and 3Department of Pediatrics, University of California–Los Angeles, Los Angeles, CA 90095, USA

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.

* To whom correspondence should be addressed at: University of California–Los Angeles, Mechanical and Aerospace Engineering Department, 420 Westwood Plaza, Room 38-137J, PO Box 951597, Los Angeles, CA 90095-1597, USA. Tel: +1 310 825 9993; Fax: +1 310 206 2302; Email: chihming{at}ucla.edu The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors


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