The Validation of Real-time PCR Assays for Infectious Diseases
from: Real-Time PCR: Advanced Technologies and Applications (Edited by: Nick A. Saunders and Martin A. Lee). Caister Academic Press, U.K. (2013)
The real-time polymerase chain reaction is now established as one of the core technologies for diagnosing infectious diseases. The early stages of the technique's development were followed by a dramatic increase in the number of diagnostic assays being published, together with the introduction of commercially produced tests. Each of the numerous publications showed a number of differences in the approach to validating the newly-produced assays and in the quality and quantity of the data supporting their validation. As a result, many workers have, at times, found it difficult to reproduce the published results from other laboratories. These difficulties can arise from e.g. a lack of information in the publication, differences in equipment between laboratories, the use of different extraction methods and sequence variations in the pathogen being detected. Over the years a number of authors have voiced their concerns over the subject of what constitutes a properly validated assay, highlighting the issues of basic scientific good practice and the responsibilities of journals in publishing full validation data. This chapter summarises the recent work covering validation and verification methodology in order to provide a practical guide to help inform and standardise the process read more ...