Guidelines for Use of External RNA Controls

The Clinical and Laboratory Standards Institute (CLSI, Wayne, PA, USA) guidelines provide protocols for the use of external RNA controls in microarray and quantitative real-time polymerase chain reaction (QRT-PCR)-based gene expression.

External RNA controls refer to RNA species that are distinct from the RNAs in the sample that is to be analyzed. The external RNA controls are added prior to enzymatic manipulations, such as reverse transcription and labeling. The protocols enable laboratory, microarray, and reagent manufacturers to assess the performance of these expression assays. They support the use of external RNA controls in microarray and QRT-PCR-based gene expression experiments.

The guidelines address important issues associated with the use of external RNA controls as a tool for verification of technical performance, and in support of the evaluation of qualitative results for a specific clinical analyte including preparation of control transcripts, design of primers and amplicons, quality control, use in final experimental or clinical test application, and the analysis and interpretation of data obtained.

Microarray and QRT-PCR technologies are emerging as vital components of genomic, evidence-based medicine and are already having an impact. Standard controls are required to ensure reliability and quality from these assay platforms before microarray and QRT-PCR results are accepted for clinical applications. The ability to report reliable gene expression results of known quality is the key to the successful employment of microarrays and QRT-PCR as diagnostic devices in clinical medicine and as tools in toxicogenomics, pharmacogenetics, and pharmacogenomics.



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