Sage Journals: Discover world-class research

Abstract

Aim: With the arrival of increasingly complex molecular tests, we are obliged to create new ways to monitor and troubleshoot the underperformance of these multiplex assays. A synthetic multiallelic quality control material has been designed to augment genomic DNA controls. We aimed to evaluate the control on a large scale, testing it on a wide variety of oligonucleotide ligation assays, test protocols, and analysis software. In addition, we investigated how laboratories treat untried and complex materials. Methods: The synthetic control monitored 32 cystic fibrosis transmembrane conductance regulator mutations and polymorphisms simultaneously. Participants of a cystic fibrosis external quality assessment scheme were invited to analyze the quality control. Results: In total, 58 laboratories participated in this study. Twenty-seven (47%) laboratories detected 32 variants; another 27 laboratories (47%) detected from 31 to 4 variants and 4 participants reported no variants (6%). The main observations included administrative errors when indicating variants on a checklist, errors caused by misreading the instructions for use of the control or assay, and technical problems related to the assay used. Conclusion: Synthetic quality control materials proved to be valuable in troubleshooting underperforming assays and complement existing genomic controls. The study also revealed a strong need for increased quality control in the postanalytical phase of testing.

Get full access to this article

View all access options for this article.

References

Bernacki

, Stankovic

, Williams

et al. 2003. Establishment of stably EBV-transformed cell lines from residual clinical blood samples for use in performance evaluation and quality assurance in molecular genetic testing. J Mol Diagn, 5:227-230.

Berwouts

, Gordon

, Rundell

et al. 2008. Evaluation and use of a synthetic quality control material, included in the European external quality assessment scheme for cystic fibrosis. Hum Mutat, 29:1063-1070.

Camajova

, Berwouts

, Matthijs

et al. 2008. Variability in the use of CE-marked assays for in vitro diagnostics of CFTR gene mutations in European genetic testing laboratories. Eur J Hum Genet, 17:537-540.

Chen

, Gagnon

, Shahangian

et al. 2009. Good laboratory practices for molecular genetic testing for heritable diseases and conditions. MMWR Recomm Rep, 58:1-37.

Chen

, O’ Connell

, Boone

et al. 2005. Developing a sustainable process to provide quality control materials for genetic testing. Genet Med, 7:534-549.

Christensen

, Jama

, Ponek

et al. 2007. Design, development, validation, and use of synthetic nucleic Acid controls for diagnostic purposes and application to cystic fibrosis testing. J Mol Diagn, 9:315-319.

Coriell Institute. 2010. NIGMS The Human Genetic Cell Repository. http://ccr.coriell.org/Sections/Collections/NIGMS/?SsId=8. 2010 December 23.

Dequeker

, Ramsden

, Grody

et al. 2001. Quality control in molecular genetic testing. Nature Rev, 2:717-723.

Devitt

, Barton

. 2008. When good CF tests go bad. Eur J Hum Genet, 17:403-405.

10.

Grody

. 2003. Quest for controls in molecular genetics. J Mol Diagn, 5:209-211.

11.

Grody

, Cutting

, Klinger

et al. 2001. Laboratory standards and guidelines for population-based cystic fibrosis carrier screening. Genet Med, 3:149-154.

12.

Hospital for Sick Children. 2010. Cystic Fibrosis Mutation Database. www.genet.sickkids.on.ca/cftr/StatisticsPage.html. 2010 December 23.

13.

Huang

, Pan

. 2007. Validation of cystic fibrosis mutation analysis using ABI 3130XL genetic analyzer. Diagn Mol Pathol, 16:57-59.

14.

International Organization for Standardization. 2007. ISO 15189:2007 Medical laboratories - Particular requirements for quality and competence.

15.

Jarvis

, Iyer

, Williams

et al. 2005. A novel method for creating artificial mutant samples for performance evaluation and quality control in clinical molecular genetics. J Mol Diagn, 7:247-251.

16.

Johnson

, Yoshitomi

, Richards

. 2007. A Comparative Study of Five Technologically Diverse CFTR Testing Platforms. J Mol Diagn, 9:401-407.

17.

Krafft

, Lichy

. 2005. Time-motion analysis of 6 cystic fibrosis mutation detection systems. Clin Chem, 51:1116-1122.

18.

Palomaki

, Haddow

, Bradley

et al. 2002. Updated assessment of cystic fibrosis mutation frequencies in non-Hispanic Caucasians. Genet Med, 4:90-94.

19.

Pratt

, Caggana

, Bridges

et al. 2009. Development of genomic reference materials for cystic fibrosis genetic testing. J Mol Diagn, 11:186-193.

20.

Richards

, Bradley

, Amos

et al. 2002. Standards and guidelines for CFTR mutation testing. Genet Med, 4:379-391.

21.

Rundell

. 2006. QC challenges for molecular testing. MLO Med Lab Obs, 38:37-41.

22.

Watson

, Cutting

, Desnick

et al. 2004. Cystic fibrosis population carrier screening: 2004 revision of American College of Medical Genetics mutation panel. Genet Med, 6:387-391.

Supplementary Material

Please find the following supplemental material available below.

For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.

For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.

0.00 MB

6.10 MB

Multiallelic Synthetic Quality Control Material: Lessons Learned from the Cystic Fibrosis External Quality Assessment Scheme

Abstract

Get full access to this article

References

Supplementary Material