HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: clinchem.2006.074807v1 53/3/531    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (3) Citing Articles via Google Scholar Google Scholar Articles by Mantovani, V. Articles by Romeo, G. Search for Related Content PubMed PubMed Citation Articles by Mantovani, V. Articles by Romeo, G. Related Collections Molecular Diagnostics and Genetics

Simple Method for Haplotyping the Poly(TG) Repeat in Individuals Carrying the IVS8 5T Allele in the CFTR Gene

¹ Medical Genetics Unit and ² Biomedical Centre Applied Research CRBA, S.Orsola-Malpighi University Hospital, Bologna, Italy; ³ Department of Experimental Evolutionary Biology, Bologna University, Bologna, Italy;

^aaddress correspondence to this author at: U.O. Genetica Medica, Policlinico S.Orsola-Malpighi, Via Massarenti, 9, 40138 Bologna, Italy; fax 39-051-6363851, e-mail mantovan{at}med.unibo.it)

Abstract

Background: The 5T allele of the polyT tract located within intron 8 of the cystic fibrosis transmembrane conductance regulator (CFTR) gene is a variant that in trans with a severe CFTR mutation can result in normal phenotype, congenital bilateral absence of vas deferens (CBAVD), or mild cystic fibrosis. The 5T allele has been associated with the skipping of exon 9, a process that seems to be influenced by an adjacent 9–13TG tandem repeat. The 12- or 13TG repeats are often associated with an abnormal phenotype. We present here a single-step method for direct haplotyping of the TG repeats in 5T carriers.

Method: The method is based on a single-step PCR, using a fluorescently labeled forward primer and a reverse allele-specific primer matching the 5T allele. We validated the test in 30 control samples of known 5T-poly(TG) haplotype and then used this method to evaluate 57 clinical samples.

Results: The expected TG genotypes were obtained for all 5T control samples, and no nonspecific amplification of either the 7T or 9T alleles was detected. In our 5T-positive collection 9 of 9 (100%) CBAVD patients, 6 of 12 (50.0%) chronic pancreatitis patients, and 12 of 36 (33.3%) individuals undergoing assisted reproduction showed 5T-12TG haplotype.

Conclusions: Our method is an accurate, specific, and simple tool to characterize the 5T poly(TG) haplotype. Our results confirm the high frequency of 5T-12TG in CBAVD patients and do not preclude a potential effect also in pancreatitis. This assay can be useful in assessment of the disease risk in 5T carriers.

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

				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]

				P.-H. Kuo, L. G. Doudeva, Y.-T. Wang, C.-K. J. Shen, and H. S. Yuan Structural insights into TDP-43 in nucleic-acid binding and domain interactions Nucleic Acids Res., April 1, 2009; 37(6): 1799 - 1808. [Abstract] [Full Text] [PDF]

				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]