Checklist for optimization and validation of real-time PCR assays.
Real-time polymerase chain reaction (PCR) is a frequently used technique in molecular diagnostics. To date, practical guidelines for the complete process of optimization and validation of commercial and in-house developed molecular diagnostic methods are scare. Therefore, we propose a practical guid...
| Publicado en: | Journal of Clinical Laboratory Analysis Vol. 23; no. 3; pp. 145 - 152 |
|---|---|
| Autores principales: | , , , , , , , |
| Formato: | research Journal Article |
| Publicado: |
Wiley-Blackwell
2009
|
| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=105541825&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 105541825 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 08878013 OCK jtl: Journal of Clinical Laboratory Analysis issn: 08878013 maglogo: Y pubinfo: dt: 2009 vid: 23 iid: 3 pid: 480 pub: Wiley-Blackwell place: Malden, Massachusetts artinfo: ui: 105541825 NLM19455629 2010298888 10.1002/jcla.20307 NLM19455629 105541825 ppf: 145 ppct: 7 formats: tig: atl: Checklist for optimization and validation of real-time PCR assays. aug: au: Raymaekers M Smets R Maes B Cartuyvels R Raymaekers, Marijke Smets, Rita Maes, Brigitte Cartuyvels, Reinoud affil: Molecular Biology, Clinical Laboratory, Virga Jesse Hospital, Hasselt, Belgium sug: subj: Cytogenetic Analysis Standards Polymerase Chain Reaction Standards Calibration Data Analysis, Statistical Genes Nucleic Acid Probes Quality Control (Technology) Reproducibility of Results Specimen Handling Human ab: Real-time polymerase chain reaction (PCR) is a frequently used technique in molecular diagnostics. To date, practical guidelines for the complete process of optimization and validation of commercial and in-house developed molecular diagnostic methods are scare. Therefore, we propose a practical guiding principle for the optimization and validation of real-time PCR assays. Based on literature, existing guidelines, and personal experience, we created a checklist that can be used in different steps of the development and validation process of commercial and in-house developed real-time PCR assays. Furthermore, determination of target values and reproducibility of internal quality controls are included, which allows a statistical follow-up of the performance of the assay. Recently, we used this checklist for the development of various qualitative and quantitative assays for microbiological and hematological applications, for which accreditation according to ISO 15189:2007 was obtained. In our experience, the use of the proposed guidelines leads to a more efficient and standardized optimization and validation. Ultimately, this results in reliable and robust molecular diagnostics. The proposed checklist is independent of environment, equipment, and specific applications and can be used in other laboratories. A worldwide consensus on this kind of checklist should be aimed at. pubtype: Academic Journal doctype: research Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
|---|