caa a22 4500 378869590 CHVBK 20180305123406.0 cr unu---uuuuu 161128e20030127xx s 000 0 eng 10.1515/CCLM.2003.006 doi (NATIONALLICENCE)gruyter-10.1515/CCLM.2003.006 Molecular Diagnosis of Cystic Fibrosis:Comparison of Four Analytical Procedures [Elektronische Daten] [Rossella Tomaiuolo, Mariacristina Spina, Giuseppe Castaldo] We compared the analytical accuracy, times, costs and the detection rate of four procedures for the molecular analysis of cystic fibrosis (CF). DNA from 127 subjects bearing different genotypes was tested by denaturating gradient gel electrophoresis followed by sequencing (reference procedure) and, for comparison, by ASO dot-blot (in-house), reverse dot-blot (Innogenetics), ARMS (Zeneca Diagnostics) and OLA-PCR (PE Applied Biosystem). To assess inter-observer variability, all samples were tested twice. To evaluate intra- and between-series variability, two samples were tested twice in each series. All the procedures yielded reproducible results and assigned the correct genotype to each sample. ASO dot-blot is the cheapest procedure but has the longest analytical time (>24 h) and uses radioactive labeling. It can be adapted to analyze peculiar mutations in specific ethnic groups. ARMS from Zeneca Diagnostics is rapid (4 h), easy to perform, but, except for the ∆F508 mutation, does not distinguish the homozygote from the heterozygote genotype. It could be used for carrier analysis in families with known mutations. OLA-PCR has the highest detection rate in most ethnic groups, is automated for capillary electrophoresis but requires a high level of operator expertise: it is suitable when collected series of samples are analyzed from large patient cohorts. Reverse dot-blot is easy to perform and can be semiautomated: it can be used as first-line screening test. Given the heterogeneity of CF mutations, commercial kits should be developed to analyze mutations peculiar to specific ethnic groups. Copyright © 2003 by Walter de Gruyter GmbH & Co. KG Medical equipment & techniques nationallicence Medical diagnosis nationallicence Diseases & disorders nationallicence Tomaiuolo Rossella aut Spina Mariacristina aut Castaldo Giuseppe aut Clinical Chemistry and Laboratory Medicine Walter de Gruyter 41/1(2003-01-27), 26-32 1434-6621 41:1<26 2003 41 cclm https://doi.org/10.1515/CCLM.2003.006 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/CCLM.2003.006 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Tomaiuolo Rossella aut NATIONALLICENCE 700 1- Spina Mariacristina aut NATIONALLICENCE 700 1- Castaldo Giuseppe aut NATIONALLICENCE 773 0- Clinical Chemistry and Laboratory Medicine Walter de Gruyter 41/1(2003-01-27), 26-32 1434-6621 41:1<26 2003 41 cclm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter