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International Research Team Publishes qPCR Guidelines

NEW YORK (GenomeWeb News) – The American Association for Clinical Chemistry and ARUP Laboratories, a national reference lab owned by the University of Utah, this week announced the publication of new consensus guidelines for researchers reporting on quantitative real-time polymerase chain reaction studies.

The "Minimum Information for Publication of Quantitative Real-Time PCR Experiments," or MIQE, guidelines appear in the April issue of the journal Clinical Chemistry. An international research team drafted the guidance document, which outlines "the minimum information necessary for evaluating qPCR experiments" and is intended to increase the transparency, reliability, quality, and reproducibility of qPCR studies.

"Currently, a lack of consensus exists on how best to perform and interpret quantitative real-time PCR (qPCR) experiments," the authors explained. "The aim of this document is to provide authors, reviewers, and editors specifications for the minimum information ... that must be reported for a qPCR experiment to ensure its relevance, accuracy, correct interpretation, and repeatability."

Since qPCR is frequently used in research studies — and diagnostic testing — the authors argued that there is a need to clarify qPCR studies are conducted, interpreted, and reported. Without such guidelines, they warned, "there is a real danger of the scientific literature being corrupted with a multitude of publications reporting inadequate and conflicting results."

Lead author Stephen Bustin, a cell and molecular biology researcher at the Queen Mary University of London, and his colleagues noted that problems such as unsuitable sample collection and storage, poor primer choices, and inappropriate data analysis can skew results.

"The majority of published qPCR studies do not provide sufficient experimental detail, and scientists reading them have a hard time deciding if the conclusions are valid," Bustin said in a statement.

In an effort to increase transparency and prevent erroneous scientific conclusions, the team outlined a checklist of conditions that qPCR studies should be required or recommended to meet before being considered for publication. The MIQE paper covers a range of qPCR-related topics — including nomenclature, conceptual tips, things to consider when using qPCR in research and diagnostic settings, sample collection and handling, nucleic acid quality control, data analysis, and more.

While it does not call for compulsory use of common reporting language, its authors note that such recommendations may be added to future updates. And though the team conceded that it may not be realistic to apply all of the criteria to large sample sets used for identifying expression signatures, they suggest the checklist is applicable once studies have been pared down to 20 targets or less.

The recommendations are modeled after Minimum Information About a Microarray Experiment (MIAME) guidelines developed for microarray studies, Minimum Information About a Proteomics Experiment (MIAPE) guidelines used for proteomics studies, and the Minimum Information About a Genome Sequence (MIGS) guidelines developed for genome sequence studies.

All of these existing efforts, as well as similar efforts aimed at standardizing RNA interference and metabolomics research, are part of the larger Minimum Information for Biological and Biomedical Investigations, or MIBBI, initiative.

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