PCR: living life amplified and standardized
With strategies for reproducibility and quality control, scientists seek to cultivate better practices using MIQE and qPCR.
Introduction to MIQE and qPCR
If MIQE, pronounced “Mikey,” were a person, he or she would be someone with a familiar-sounding name who is held in high regard but is not close to many. MIQE urgently wants more friends.
MIQE stands for Minimum Information for Publication of Quantitative Real-Time PCR Experiments, which is a set of guidelines published in 2009 by scientists from 12 academic institutions and 2 companies1, and it has been cited nearly 1,300 times since publication. Despite the guidelines and despite the long tenure of quantitative polymerase chain reaction (qPCR, also known as real-time PCR) as a detection and quantification tool, the challenges of this technique seem to fall smack into many scientists’ blind spot.
Changing that reality takes nagging and appeals, practical recommendations and new approaches—all in the name of achieving more reliable, reproducible results from qPCR experiments that will shape the backbone of future clinical diagnostic tests. Despite the critical consequences of irreproducible results (Box 1), workshop organizers see both good and problematic behavior in labs using PCR (Box 2). Quality-control issues in qPCR are moving to the fore through activities by the MIQE authors and initiatives such as the European Union’s public-private venture SPIDIA: Standardisation and improvement of generic pre-analytical tools and procedures for in vitro diagnostics. A recent SPIDIA paper assessing the practices in over 100 labs across Europe relating to the quality of RNA extracted from blood samples revealed a wide disparity in extraction procedures and processing. Around 30% of the labs had at least two problematic quality-control parameters2.
