Abstract
Background: The Italian External Quality Control Programme for cystic fibrosis molecular diagnosis started in 2001; public laboratories distributed throughout Italy participated on a voluntary basis.
Methods: The Italian Public Health Institute (Istituto Superiore di Sanità) sent six validated DNA samples to participating laboratories: technical and clinical information was provided for each sample. Laboratories were required to analyse all six samples. For each sample the laboratories had to provide the results (including raw data) and a report of molecular analysis within 2 months using current methods and nomenclature. Raw data and reports were evaluated by a Steering Committee and their comments were sent to each laboratory.
Results: Genotyping results indicated a general good level of quality for all laboratories, i.e., ∼1% of alleles were incorrectly assigned each year due to analytical (45%) and misinterpretation (45%) errors. During the first 2 years, more than 70% of laboratories did not test for some regional Italian mutations. Commercial kits for reverse dot-blot and oligonucleotide ligation assay PCR were used to detect mutations by 52.8% and 29.5%, respectively, of the participating laboratories. Reporting of results was still inadequate; in 2004 a model for the written report was introduced, but not all laboratories used it.
Conclusions: Our data show that few genotyping errors were made by laboratories and were principally due to misinterpretation and analytical reasons. However, reports are still inadequate and it will be interesting to evaluate the introduction of the reporting model in future years.
Clin Chem Lab Med 2007;45:254–60.
References
1. Dugueperoux I, De Braekeleer M, the Participating Centres to the French National Cystic Fibrosis Registry. Genotype-phenotype relationship for five CFTR mutations frequently identified in western France. J Cyst Fibros2004;3:259–63.10.1016/j.jcf.2004.07.004Search in Google Scholar
2. Castaldo G, Polizzi A, Tomaiuolo R, Cazeneuve C, Girodon E, Santostasi T, et al. Comprehensive cystic fibrosis mutation epidemiology and haplotype characterization in a southern Italian population. Ann Hum Genet2005;69:15–24.10.1046/j.1529-8817.2004.00130.xSearch in Google Scholar
3. Estivill X, Bancells C, Ramos C. The Biomed CF Mutation Analysis Consortium. Geographic distribution and regional origin of 272 cystic fibrosis mutations in European populations. Hum Mutat1997;10:135–54.10.1002/(SICI)1098-1004(1997)10:2<135::AID-HUMU6>3.0.CO;2-JSearch in Google Scholar
4. The CF mutation database. http://www.genet.sickkids.on.ca/cftr. Accessed January 15, 2007.Search in Google Scholar
5. Kerem B, Rommens JM, Buchanan JA, Markiewicz D, Cox TK, Chakravarti A, et al. Identification of the cystic fibrosis gene: genetic analysis. Science1989;245:1073–80.10.1126/science.2570460Search in Google Scholar
6. Bobadilla JL, Macek M Jr, Fine JP, Farrell PM. Cystic fibrosis: a worldwide analysis of CFTR mutations – correlation with incidence data and application to screening. Hum Mutat2002;19:575–606.10.1002/humu.10041Search in Google Scholar
7. Bombieri C, Pignatti PF. Cystic fibrosis mutation testing in Italy. Genet Test2001;5:229–33.10.1089/10906570152742281Search in Google Scholar
8. Ferec C, Le Marechal C, Audrezet MP, Farinha CM, Amaral MD, Gallati S, et al. Analysis of genomic CFTR DNA. J Cyst Fibros2004;3:7–10.10.1016/j.jcf.2004.05.003Search in Google Scholar
9. Chevalier-Porst F, Souche G, Bozon D. Identification and characterization of three large deletions and deletion/polymorphism in the CFTR gene. Hum Mutat2005;25:504.10.1002/humu.9335Search in Google Scholar
10. Castellani C, Bonizzato A, Cabrini G, Mastella G. Newborn screening strategy for cystic fibrosis: a field study in an area with high allelic heterogeneity. Acta Paediatr1997;86:497–502.10.1111/j.1651-2227.1997.tb08920.xSearch in Google Scholar
11. Castellani C, Picci L, Scarpa M, Dechecchi MC, Zanolla L, Assael BM, et al. Cystic fibrosis carriers have higher neonatal immunoreactive trypsinogen values than non-carriers. Am J Med Genet2005;135A:142–4.10.1002/ajmg.a.30470Search in Google Scholar
12. Foresta C, Ferlin A, Gianaroli L, Dallapiccola B. Guidelines for the appropriate use of genetic tests in infertile couples. Eur J Hum Genet2002;10:303–12.10.1038/sj.ejhg.5200805Search in Google Scholar PubMed
13. Dequeker E, Cassiman JJ. Genetic testing and quality control in diagnostic laboratories. Nat Genet2000;25:259–60.10.1038/77008Search in Google Scholar PubMed
14. Dequeker E, Ramsden S, Grody WW, Stenzel TT, Barton DE. Quality control in molecular genetic testing. Nat Rev Genet2001;2:717–23.10.1038/35088588Search in Google Scholar PubMed
15. Holtzman N, Shapiro D. Genetic testing and public policy. Br Med J1998;316:852–6.10.1136/bmj.316.7134.852Search in Google Scholar PubMed PubMed Central
16. Schwartz M. Genetic testing and the clinical laboratory improvement amendments of 1988: present and future. Clin Chem1999;45:739–45.10.1093/clinchem/45.5.739Search in Google Scholar
17. The Italian National Centre for Rare Diseases. http://www.iss.it/cnmr. Accessed January 15, 2007.Search in Google Scholar
18. Taruscio D, Falbo V, Floridia G, Salvatore M, Pescucci C, Cantafora A, et al. Quality assessment in cytogenetic and molecular genetic testing: the experience of the Italian Project on Standardisation and Quality Assurance. Clin Chem Lab Med2004;42:915–21.10.1515/CCLM.2004.148Search in Google Scholar PubMed
19. Rendine S, Calafell F, Cappello N, Gagliardini R, Caramia G, Rigillo N, et al. Genetic history of cystic fibrosis mutations in Italy. I. Regional distribution. Ann Hum Genet1997;61:411–24.10.1017/S0003480097006477Search in Google Scholar
20. Database of Italian laboratories performing genetic testing for CF mutation. http://spazioinwind.libero.it/laboratoricf/. Accessed January 15, 2007.Search in Google Scholar
21. Burke W, Zimmern RL. Ensuring the appropriate use of genetic tests. Nat Rev Genet2004;5:955–9.10.1038/nrg1495Search in Google Scholar PubMed
22. Batjer JD. The College of American Pathologists laboratory accreditation program. Clin Lab Haematol1990;12:135–8.Search in Google Scholar
23. Dequeker E, Cuppens H, Dodge J, Estivill X, Goossens M, Pignatti PF, et al. Recommendations for quality improvement in genetic testing for cystic fibrosis. European Concerted Action on Cystic Fibrosis. Eur J Hum Genet2000;8:S2–24.10.1038/sj.ejhg.5200487Search in Google Scholar PubMed
24. Van Loon C, Van Crombruggen J, Azzuz I, Celis L, Bläser M, Timmermans L, et al. Validation of a new line probe assay for the amplification and simultaneous detection of 21 Italian CFTR mutations. Presented at the 28th European Cystic Fibrosis Conference, Crete, June 22–25, 2005.Search in Google Scholar
25. Lemna WK, Feldman GL, Kerem B, Fernbach SD, Zevkovich EP, O'Brien WE, et al. Mutation analysis for heterozygote detection and the prenatal diagnosis of cystic fibrosis. N Engl J Med1990;322:291–6.10.1056/NEJM199002013220503Search in Google Scholar PubMed
26. Castellani C, Lalatta F, Neri D, Novelli G, Piccinini A, Seia M, et al. Modelli di analisi genetica per fibrosi cistica a cura della Commissione di Studio sulle modalità di analisi genetica per Fibrosi Cistica. http://www.pnlg.it/out/005/005.pdf. Accessed January 15, 2007.Search in Google Scholar
©2007 by Walter de Gruyter Berlin New York
