Analysis of 31 CFTR mutations by polymerase chain reaction/oligonucleotide ligation assay in a pilot screening of 4476 newborns for cystic fibrosis

Abstract

Objectives-Molecular biological testing for genetic diseases has grown rapidly, but speed, accuracy, specificity, sensitivity, throughput, and cost become more important as large scale screening is considered. This is a pilot study of an assay for the simultaneous detection of up to 31 cystic fibrosis mutations in a multicentre population based screening of 4476 Italian newborns. Conclusions-PCR/OLA is a robust, accurate, user friendly method for cystic fibrosis screening of newborns using blood spots in a semiautomated way at a low cost per mutation (0.8 Euro). (J Med Screen 1999;6:67-69) Methods-The

Mutation analysis of the cystic fibrosis transmembrane conductance regulator (CFTR) gene, the cationic trypsinogen (PRSS1) gene, and the serine protease inhibitor, Kazal type 1 (SPINK1) gene in patients with alcoholic chronic pancreatitis

“…Other seven mutations, frequently observed in Italian CF patients (Q552X; 711+5G4A; 2790-2A4G; S589N; T338I; 1898+3A4G, and 1717-8G4A), were examined by sequencing. Using both methods in the general population from the same geographical area (San Giovanni Rotondo) in which our study was performed, Gasparini et al 23 found a carrier frequency of CFTR mutations of 3.22%. The CFTR polymorphic intron 8 polyT region was analyzed according to the method described by Chillon et al 24 This sequence contains five, seven, or nine thymidines (5T, 7T, 9T, respectively).…”

Negative sweat test in hypertrypsinaemic infants with cystic fibrosis carrying rare CFTR mutations

“…Genetic analysis was initially performed by PCR and oligonucleotide ligation assay (OLA) [10], then an extended analysis of the CFTR gene was performed using denaturing gradient gel electrophoresis (DGGE) analysis and sequencing as previously reported [1]. An allele-specific PCR assay was used to distinguish the 5T, 7T and 9T alleles.…”

Potential role for the common cystic fibrosis ΔF508 mutation in Crohnʼs disease

“…As shown in Table 2, the frequency of ⌬F508 did not significantly differ between UC patients and controls, and was comparable to that expected for a general population from Italy (2.1%). 28 On the contrary, this mutation was underrepresented in the CD group, where it occurred at a 8-fold lower frequency compared with controls (P ϭ 0.021). Accordingly, heterozygous carriers had an estimated 88% reduced risk of developing CD in this population (OR ϭ 0.12).…”