Back to Search
Start Over
EQUAL-quant: an international external quality assessment scheme for real-time PCR
- Source :
- Clinical chemistry. 52(8)
- Publication Year :
- 2006
-
Abstract
- Background: Quantitative gene expression analysis by real-time PCR is important in several diagnostic areas, such as the detection of minimum residual disease in leukemia and the prognostic assessment of cancer patients. To address quality assurance in this technically challenging area, the European Union (EU) has funded the EQUAL project to develop methodologic external quality assessment (EQA) relevant to diagnostic and research laboratories among the EU member states. We report here the results of the EQUAL-quant program, which assesses standards in the use of TaqMan™ probes, one of the most widely used assays in the implementation of real-time PCR. Methods: The EQUAL-quant reagent set was developed to assess the technical execution of a standard TaqMan assay, including RNA extraction, reverse transcription, and real-time PCR quantification of target DNA copy number. Results: The multidisciplinary EQA scheme included 137 participating laboratories from 29 countries. We demonstrated significant differences in performance among laboratories, with 20% of laboratories reporting at least one result lacking in precision and/or accuracy according to the statistical procedures described. No differences in performance were observed for the >10 different testing platforms used by the study participants. Conclusions: This EQA scheme demonstrated both the requirement and demand for external assessment of technical standards in real-time PCR. The reagent design and the statistical tools developed within this project will provide a benchmark for defining acceptable working standards in this emerging technology.
- Subjects :
- Scheme (programming language)
Quality Control
Pathology
medicine.medical_specialty
Clinical Biochemistry
Technical standard
Gene Expression
Polymerase Chain Reaction
External quality assessment
medicine
TaqMan
media_common.cataloged_instance
Humans
Medical physics
European Union
European union
Proto-Oncogene Proteins c-abl
computer.programming_language
media_common
business.industry
Member states
Biochemistry (medical)
Real-time polymerase chain reaction
Molecular Diagnostic Techniques
business
Laboratories
Quality assurance
computer
Subjects
Details
- ISSN :
- 00099147
- Volume :
- 52
- Issue :
- 8
- Database :
- OpenAIRE
- Journal :
- Clinical chemistry
- Accession number :
- edsair.doi.dedup.....facc5f8d59e605da338217bd0037c701