| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
1 Department of Pediatrics, The Children’s Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104.
2 Unita’ di Ricerca in Aterosclerosi e Trombosi, IRCCS Casa Sollievo della Sofferenza, 71013 San Giovanni Rotondo, Italy.
3 Dipartimento di Science Neurologiche, Ospedale Maggiore Policlinico, University of Milan, 20122 Milan, Italy.
4 Laboratorio di Genetica Molecolare, Istituto G. Gaslini, 16148 Genova, Italy.
5 Unità di Genomica per la Diagnostica delle Patologie Umane, IRCCS H. San Raffaele, Diagnostica e Ricerca San Raffaele S.p.A., 20132 Milan, Italy.
6 Dipartimento di Medicina Sperimentale e Patologia, Università "La Sapienza", 00198 Roma; IRCCS–C.S.S. San Giovanni Rotondo and C.S.S.–Mendel, 00198 Rome, Italy.
7 Laboratorio di Biologia Molecolare, Ospedale Infantile Regina Margherita, 10124 Torino, Italy.
8 Department of Medicine, Cardeza Foundation for Hematologic Research, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107.
aAddress correspondence to this author at: Thomas Jefferson University, Medical Office Bldg, Room 406, 1100 Walnut St., Philadelphia, PA 19107. Fax 215-503-2803; e-mail paolo.fortina{at}mail.tju.edu.
Background: Microelectronic DNA chip devices represent an emerging technology for genotyping. We developed methods for detection of single-nucleotide polymorphisms (SNPs) in clinically relevant genes.
Methods: Primer pairs, with one containing a 5'-biotin group, were used to PCR-amplify the region encompassing the SNP to be interrogated. After denaturation, the biotinylated strand was electronically targeted to discrete sites on streptavidin-coated gel pads surfaces by use of a Nanogen Molecular Workstation. Allele-specific dye-labeled oligonucleotide reporters were used for detection of wild-type and variant sequences. Methods were developed for SNPs in genes, including factor VII, ß-globin, and the RET protooncogene. We genotyped 331 samples for five DNA variations in the factor VII gene, >600 samples from patients with ß-thalassemia, and 15 samples for mutations within the RET protooncogene. All samples were previously typed by various methods, including DNA sequence analysis, allele-specific PCR, and/or restriction enzyme digestion of PCR products.
Results: Analysis of amplified DNA required 4–6 h. After mismatched DNA was removed, signal-to-noise ratios were >5. More than 940 samples were typed with the microelectronic array platform, and results were totally concordant with results obtained previously by other genotyping methods.
Conclusions: The described protocols detect SNPs of clinical interest with results comparable to those of other genotyping methods.
The following articles in journals at HighWire Press have cited this article:
|
|
 |
|
  S. Galbiati, B. Foglieni, M. Travi, C. Curcio, G. Restagno, L. Sbaiz, M. Smid, F. Pasi, A. Ferrari, M. Ferrari, et al. Peptide-nucleic acid-mediated enriched polymerase chain reaction as a key point for non-invasive prenatal diagnosis of {beta}-thalassemia Haematologica, April 1, 2008; 93(4): 610 - 614. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Stenirri, G. Restagno, G. B. Ferrero, G. Alaimo, L. Sbaiz, C. Mari, L. Genitori, F. Maurizio, and L. Cremonesi Integrated Strategy for Fast and Automated Molecular Characterization of Genes Involved in Craniosynostosis Clin. Chem., October 1, 2007; 53(10): 1767 - 1774. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Glynou, P. Kastanis, S. Boukouvala, V. Tsaoussis, P. C. Ioannou, T. K. Christopoulos, J. Traeger-Synodinos, and E. Kanavakis High-Throughput Microtiter Well-Based Chemiluminometric Genotyping of 15 HBB Gene Mutations in a Dry-Reagent Format Clin. Chem., March 1, 2007; 53(3): 384 - 391. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. GALBIATI, G. RESTAGNO, B. FOGLIENI, S. BONALUMI, M. TRAVI, A. PIGA, L. SBAIZ, M. CHIARI, F. DAMIN, M. SMID, et al. Different Approaches for Noninvasive Prenatal Diagnosis of Genetic Diseases Based on PNA-Mediated Enriched PCR. Ann. N.Y. Acad. Sci., September 1, 2006; 1075: 137 - 143. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. A. Saunders, S. Alexander, and I. Tatt env Gene Typing of Human Immunodeficiency Virus Type 1 Strains on Electronic Microarrays J. Clin. Microbiol., April 1, 2005; 43(4): 1910 - 1916. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Maekawa, T. Nagaoka, T. Taniguchi, H. Higashi, H. Sugimura, K. Sugano, H. Yonekawa, T. Satoh, T. Horii, N. Shirai, et al. Three-Dimensional Microarray Compared with PCR-Single-Strand Conformation Polymorphism Analysis/DNA Sequencing for Mutation Analysis of K-ras Codons 12 and 13 Clin. Chem., August 1, 2004; 50(8): 1322 - 1327. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Frusconi, B. Giusti, L. Rossi, S. Bernabini, F. Poggi, I. Giotti, R. Abbate, G. Pepe, and F. Torricelli Improvement of Low-Density Microelectronic Array Technology to Characterize 14 Mutations/Single-Nucleotide Polymorphisms from Several Human Genes on a Large Scale Clin. Chem., April 1, 2004; 50(4): 775 - 777. [Full Text] [PDF]
|
|
|
|
|
|
B. Foglieni, L. Cremonesi, M. Travi, A. Ravani, A. Giambona, M. C. Rosatelli, C. Perra, P. Fortina, and M. Ferrari {beta}-Thalassemia Microelectronic Chip: A Fast and Accurate Method for Mutation Detection Clin. Chem., January 1, 2004; 50(1): 73 - 79. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Erali, B. Schmidt, E. Lyon, and C. Wittwer Evaluation of Electronic Microarrays for Genotyping Factor V, Factor II, and MTHFR Clin. Chem., May 1, 2003; 49(5): 732 - 739. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
