Clinical Chemistry
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 52: 361-369, 2006. First published January 26, 2006; 10.1373/clinchem.2005.061192
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Data Supplements
Right arrow All Versions of this Article:
clinchem.2005.061192v1
52/3/361    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Web of Science (1)
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Kao, H.-Y.
Right arrow Articles by Chen, Y.-J.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Kao, H.-Y.
Right arrow Articles by Chen, Y.-J.
Related Collections
Right arrow Molecular Diagnostics and Genetics
Right arrow General Clinical Chemistry
Right arrow Pediatric Clinical Chemistry
Right arrow Automation and Analytical Techniques
(Clinical Chemistry. 2006;52:361-369.)
© 2006 American Association for Clinical Chemistry, Inc.

Molecular Diagnostics and Genetics

Determination of SMN1/SMN2 Gene Dosage by a Quantitative Genotyping Platform Combining Capillary Electrophoresis and MALDI-TOF Mass Spectrometry

Hung-Yi Kao1,2,2, Yi-Ning Su3,2, Hsin-Kai Liao1, Ming S. Liu4 and Yu-Ju Chen1,5,a

1 Institute of Chemistry, and Genomics Research Center, Academia Sinica, Taipei, Taiwan, ROC.
2 Department of Bioinformatics and Structure Biology, National Tsing Hua University, Hsinchu, Taiwan, ROC.
3 Department of Medical Genetics, National Taiwan University Hospital, Taipei, Taiwan, ROC.
4 Institute eGene, Inc., Irvine, CA.
5 Institute of Molecular Biology, National Chung-Hsing University, Taichun, Taiwan.

aAddress correspondence to this author at: Institute of Chemistry, Academia Sinica, Taipei 11529, Taiwan, ROC. Fax 886-2-2783-1237; e-mail yjchen{at}chem.sinica.edu.tw.

Background: Spinal muscular atrophy (SMA) is a common inherited and fatal neuromuscular disease caused by deletions and/or mutations that lead to altered concentrations of proteins encoded by the survival motor neuron genes SMN1 and SMN2. Because of the high incidence (at least 1 in 10 000 live births and a carrier frequency of 1 in 35 to 1 in 50) and severity of the disease, precise quantification of SMN1 and SMN2 gene copy numbers is essential for diagnosis and genetic counseling.

Methods: We developed a genotyping platform combining capillary electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to quantify absolute gene dosage. The absolute gene dosage can be determined by a multiplexed competitive PCR protocol followed by capillary electrophoresis analysis. The relative SMN1/SMN2 ratio can be analyzed by PinPoint assay followed by MALDI-TOF MS analysis.

Results: The complementary assays were evaluated in confirmed cases including 9 affected patients, 33 carriers, and 478 healthy individuals from the general population. We were able to determine all genotypes with different SMN1/SMN2 gene copy number ratios, which unambiguously diagnosed carrier status and the severity of SMA with 100% specificity.

Conclusions: This quantitative genotyping platform is suitable for detection of SMA. The described approach may serve as a general quantitative genotyping method for molecular diagnosis of other inheritable diseases.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2006 by the American Association for Clinical Chemistry.