Document Detail


Certification of reference materials for detection of the human prothrombin gene G20210A sequence variant.
MedLine Citation:
PMID:  18298350     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
BACKGROUND: There is a need for reference materials (RMs) in the field of genetic testing for verification of test results obtained in patients and probands. For the frequent genetic variation G20210A in the prothrombin gene, it has been shown that purified plasmids containing the gene fragment harbouring the mutation constitute good candidate RMs. METHODS: Plasmid-type RMs were characterised for homogeneity, stability, sequence identity and fitness for purpose. Their certification required the use of different real-time PCR methods for genotyping and quantification of the plasmid copy number. RESULTS: Homogeneity, stability and fitness for the purpose of the plasmids could be demonstrated. The long-term stability (up to 24 months) of the materials was confirmed by highly sensitive and specific quantitative real-time PCR methods. CONCLUSIONS: New types of certified RMs (CRMs) for genetic testing of the human prothrombin gene G20210A sequence variant are available. Their fitness for purpose was demonstrated and no evidence was found that they would not work with other methods as long as these are targeting the whole or parts of the prothrombin gene fragment inserted into the plasmids. The described CRMs support the efforts of the international community in development, validation and harmonisation of tests for molecular genetic testing.
Authors:
David Gancberg; Philippe Corbisier; Nele Meeus; Janos Marki-Zay; Christine Mannhalter; Heinz Schimmel
Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Clinical chemistry and laboratory medicine : CCLM / FESCC     Volume:  46     ISSN:  1434-6621     ISO Abbreviation:  Clin. Chem. Lab. Med.     Publication Date:  2008  
Date Detail:
Created Date:  2008-07-09     Completed Date:  2008-07-31     Revised Date:  2008-11-21    
Medline Journal Info:
Nlm Unique ID:  9806306     Medline TA:  Clin Chem Lab Med     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  463-9     Citation Subset:  IM    
Affiliation:
European Commission, Joint Research Centre, Institute for Reference Materials and Measurements, Geel, Belgium. david.gancberg@ec.europa.eu
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MeSH Terms
Descriptor/Qualifier:
Alleles
Blood Chemical Analysis / methods,  standards*
Calibration
Genetic Variation
Genotype
Humans
Models, Genetic
Organic Chemicals / pharmacology
Plasmids / metabolism
Polymerase Chain Reaction
Prothrombin / genetics*
Reference Standards
Reference Values
Reverse Transcriptase Polymerase Chain Reaction
Temperature
Chemical
Reg. No./Substance:
0/Organic Chemicals; 0/PicoGreen; 9001-26-7/Prothrombin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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