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


     

Clinical Chemistry 0: clinchem.2005.052159v1, 2005; 10.1373/clinchem.2005.052159
This Article
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
clinchem.2005.052159v1
51/9/1619    most recent
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
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 HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Millson, A.
Right arrow Articles by Lyon, E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Millson, A.
Right arrow Articles by Lyon, E.

Received on April 12, 2005
Accepted on June 24, 2005

Molecular Diagnostics and Genetics

Direct Molecular Haplotyping of the IVS-8 Poly(TG) and PolyT Repeat Tracts in the Cystic Fibrosis Gene by Melting Curve Analysis of Hybridization Probes

Alison Millson 1, Genevieve Pont-Kingdon 1, Sam Page 1, Elaine Lyon 2

1 Institute for Clinical and Experimental Pathology, ARUP Laboratories, Salt Lake City, Utah
2 Institute for Clinical and Experimental Pathology, ARUP Laboratories, Salt Lake City, Utah, and Department of Pathology, University of Utah School of Medicine, Salt Lake City, Utah, and Address correspondence to this author at: ARUP Laboratories, 500 Chipeta Way, Salt Lake City, UT 84108

Background: Molecular haplotyping is a developing technology with great potential for use in clinical diagnostics. We describe a haplotyping method that uses PCR combined with hybridization probes.

Methods: We designed a LightCycler assay that uses fluorescence resonance energy transfer hybridization probes to haplotype the poly(TG) and polyT (TG-T) tract in the IVS-8 region of the CFTR gene. The reporter probe was designed as a perfect match to the TG12-5T allele.

Results: Analysis of 132 samples revealed 9 unique derivative melting temperatures (Tms); the lowest was 42.4 °C and the highest was 63.6 °C. The lowest Tms were in the TGn-9T group, the intermediate Tms in the TGn-7T group, and the highest Tms in the TGn-5T group. Haplotype frequencies were highest (39%) for TG11-7T and lowest (0.4%) for TG13-5T.

Conclusions: Different combinations of polymorphisms under the reporter hybridization probe had unique and characteristic Tms. This property enables genotyping as well as determination of the phase of multiple variants under the probe, a principle we demonstrated by haplotyping the TG-T repeat tract in the IVS-8 region of the CFTR gene.


The following articles in journals at HighWire Press have cited this article:


Home page
 Clin. Chem.Home page
J. K. Bickmann, W. Kamin, M. Wiebel, F. Hauser, J. J. Wenzel, C. Neukirch, M. Stuhrmann, K. J. Lackner, and H. Rossmann
A Novel Approach to CFTR Mutation Testing by Pyrosequencing-Based Assay Panels Adapted to Ethnicities
Clin. Chem., June 1, 2009; 55(6): 1083 - 1091.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
C. Costa, J.-M. Costa, J. Martin, B. Boissier, M. Goossens, and E. Girodon
Multiplex Allele-Specific Fluorescent PCR for Haplotyping the IVS8 (TG)m(T)n Locus in the CFTR Gene
Clin. Chem., September 1, 2008; 54(9): 1564 - 1567.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
G. Pont-Kingdon, R. L. Margraf, K. Sumner, A. Millson, E. Lyon, and E. Schutz
Design and Application of Noncontinuously Binding Probes Used for Haplotyping and Genotyping
Clin. Chem., June 1, 2008; 54(6): 990 - 999.
[Abstract] [Full Text] [PDF]

Home page
 J. Mol. Diagn.Home page
C. Bareil, C. Guittard, J.-P. Altieri, C. Templin, M. Claustres, and M. des Georges
Comprehensive and Rapid Genotyping of Mutations and Haplotypes in Congenital Bilateral Absence of the Vas Deferens and Other Cystic Fibrosis Transmembrane Conductance Regulator-Related Disorders
J. Mol. Diagn., November 1, 2007; 9(5): 582 - 588.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
V. Mantovani, P. Garagnani, P. Selva, C. Rossi, S. Ferrari, M. Cenci, N. Calza, V. Cerreta, D. Luiselli, and G. Romeo
Simple Method for Haplotyping the Poly(TG) Repeat in Individuals Carrying the IVS8 5T Allele in the CFTR Gene
Clin. Chem., March 1, 2007; 53(3): 531 - 533.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
B. A. Konfortov, A. T. Bankier, and P. H. Dear
An efficient method for multi-locus molecular haplotyping
Nucleic Acids Res., January 12, 2007; 35(1): e6 - e6.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
C. Costa, M. Goossens, and E. Girodon
Simultaneous Molecular Haplotyping of Both IVS8 (TG)m and (T)n Tracts in the CFTR Gene: Still a Challenge.
Clin. Chem., August 1, 2006; 52(8): 1621 - 1622.
[Full Text] [PDF]

Home page
 Clin. Chem.Home page
A. Millson, G. Pont-Kingdon, and E. Lyon
Response to the letter "Simultaneous Molecular Haplotyping of Both IVS8 (TG)m and (T)n Tracts in the CFTR Gene: Still a Challenge" by Costa et al.
Clin. Chem., August 1, 2006; 52(8): 1622 - 1622.
[Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH
Copyright © 2005 by the American Association for Clinical Chemistry.