**5. The MIQE guidelines**

The problems inherent in the design and reporting of qPCR-based assays had been known for a long time, but there had been no concerted effort to tackle this serious problem. The 2002 O'Leary paper is not unique in omitting sufficient experimental detail, which impedes the readers' ability to evaluate critically the quality of the results presented, repeat the re‐ ported experiments, or integrate methodological advances into their own studies. Indeed, a recent survey of qPCR-based publication demonstrates very clearly that this problem per‐ sists even in 2010 [20] and that the perceived quality of the publishing journal is not correlat‐ ed with the actual quality of the qPCR publication. The egregious use of qPCR in the MeV/MMR/autism context provided the stimulus to initiate a push towards improving the technical aspects of qPCR assay design and reporting. Consequently, guidelines tackling this issue have recently been published which aim to promote consistency between laboratories, and increase experimental transparency [21]. The Minimum Information for publication of Quantitative real-time PCR Experiments (MIQE) guidelines outline the minimum data set necessary to evaluate effectively RT-qPCR assays, and are designed to be used as a checklist to both accompany manuscript submission and to be available alongside the published manuscript to enhance critical appraisal.

The four key areas of standardisation that define any qPCR experiment are study design, technical detail, analysis methods and statistics. MIQE addresses these under a set of cap‐ tions that describe a large number of individual elements: "Experimental design, sample, nucleic acids, reverse transcription, target, primers and probes, assay details, PCR cycling and data analysis". At first sight, these look daunting, arduous and over-exacting. In prac‐ tice, it is clear that most, if not all of these parameters describe information that would be obtained as a matter of course during the experimental design, optimisation and validation stages. Importantly, there is a clear hierarchy with some parameters, labelled "E" (essential) in the published guidelines, indispensable for an adequate description of the qPCR assay, whereas other components, labelled "D" (desirable) more peripheral, yet constituting an ef‐ fective foundation for the realisation of best practice protocols. There is increasing recogni‐ tion that the MIQE guidelines provide the basis for much-needed standardisation as well as encourage the publication of essential information that should be accessible to reviewer and reader[22].

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