Detection of <b><i>CFTR</i></b> mutations using temporal temperature gradient gel electrophoresis

Wong, Lee‐Jun C.; Alper, Özgül M.

doi:10.1002/elps.200406015

Cited by 14 publications

(8 citation statements)

References 31 publications

(51 reference statements)

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“…10,13,14,20,29,30 Differences in detected mutations among these studies may be due in part to the fact that the Hispanic population is not homogeneous and is typically defined more by geographic origins in various regions of southern Europe or the Americas, rather than by one specific genetic background. 12 Overall, mutation detection in the CFTR gene has largely focused on point mutations, small deletions and small insertions within or close to exons.…”

Section: Discussionmentioning

confidence: 99%

Multiplex Ligation-Dependent Probe Amplification Identification of Whole Exon and Single Nucleotide Deletions in the CFTR Gene of Hispanic Individuals with Cystic Fibrosis

Schrijver

Rappahahn

Pique

et al. 2008

The Journal of Molecular Diagnostics

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A disparity between Caucasian and Hispanic mutation detection for cystic fibrosis continues to exist, although the carrier frequency is only moderately lower in Hispanics. We aimed to identify exonic rearrangements that remained undetected by conventional methods. In seven of 32 cystic fibrosis-affected self-identified Hispanics for whom only one or no mutations were identified by extensive molecular testing, exon deletions appeared to be present with a multiplex ligation-dependent probe amplification (MLPA) assay. Two recurrent deletions (of exons 2-3 and exons 22-23) were identified in one and three patients, respectively (12.5%, 11.1% of unidentified alleles). Two apparently novel deletions (exons 6b and 20) were identified in three additional patients. Subsequent sequencing to characterize deletion breakpoints, however, identified single nucleotide deletions at the probe binding sites close to the ligation point. All resulted in false positive MLPA deletion signals. Interestingly, these mutations were not common in Caucasians, and one (935delA) was common in U.S. Hispanics. On examination of all probe binding sites, we identified a total of 76 reported mutations and five silent variants that immediately surrounded the MLPA ligation sites, with 22 occurring in non-Caucasians. These mutations are not all rare. Thus, apparent exon deletions by MLPA may indicate the presence of both large deletions and point mutations, with important implications for pan-ethnic MLPA testing in cystic fibrosis and other genetic conditions. (J Mol

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Section: Discussionmentioning

confidence: 99%

Multiplex Ligation-Dependent Probe Amplification Identification of Whole Exon and Single Nucleotide Deletions in the CFTR Gene of Hispanic Individuals with Cystic Fibrosis

Schrijver

Rappahahn

Pique

et al. 2008

The Journal of Molecular Diagnostics

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“…To be certain that no patients will be missed, a DGGE analysis or similar technology following the OLA analysis is necessary, as is shown by the patient carrying a ^F508 mutation and the E60X mutation that was discovered through the DGGE analysis. Although we used DGGE analysis, currently other rapid and sensitive mutation analysis techniques are available, such as temperature gradient capillary electrophoresis (TGCE) [10] or temporal temperature gradient gel analysis (TGGE) [11], which are probably even more efficient in CFTR mutation analysis.…”

Section: Discussionmentioning

confidence: 99%

Extended gene analysis can increase specificity of neonatal screening for cystic fibrosis

et al. 2006

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“…TTGE has been shown to be a sensitive screening tool for detecting novel mutations in the human CFTR gene (27). This technique can detect single base substitutions, insertions, and deletions.…”

Section: Methodsmentioning

confidence: 99%

Naturally occurring mutations in the canine CFTR gene

Spadafora

Hawkins

Murphy

et al. 2010

Physiological Genomics

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Naturally occurring cystic fibrosis (CF)-causing mutations in the CFTR gene have not been identified in any nonhuman animal species. Since domestic dogs are known to develop medical conditions associated with atypical CF in humans (e.g., bronchiectasis and pancreatitis), we hypothesized that dogs with these disorders likely have a higher expression rate of CFTR mutations than the at-large population. Temporal temperature-gradient gel electrophoresis (TTGE) was used to screen canine CFTR in 400 animals: 203 dogs diagnosed with pancreatitis, 23 dogs diagnosed with bronchiectasis, and 174 dogs admitted to clinics for any illness (at-large dogs). Twenty-eight dogs were identified with one of four CFTR missense mutations. P1281T and P1464H mutations occur in relatively unconserved residues. R1456W is analogous to the human R1453W mutation, which has ∼20% of normal CFTR function and is associated with pancreatitis and panbronchiolitis. R812W disrupts a highly conserved protein kinase A recognition site within the regulatory domain. We conclude that naturally occurring CFTR mutations are relatively common in domestic dogs and can be detected with TTGE. No substantive differences in mutation frequency were observed between the at-large, pancreatitis, and bronchiectasis dogs.

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Detection of CFTR mutations using temporal temperature gradient gel electrophoresis

Cited by 14 publications

References 31 publications

Multiplex Ligation-Dependent Probe Amplification Identification of Whole Exon and Single Nucleotide Deletions in the CFTR Gene of Hispanic Individuals with Cystic Fibrosis

Multiplex Ligation-Dependent Probe Amplification Identification of Whole Exon and Single Nucleotide Deletions in the CFTR Gene of Hispanic Individuals with Cystic Fibrosis

Extended gene analysis can increase specificity of neonatal screening for cystic fibrosis

Naturally occurring mutations in the canine CFTR gene

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