We’ve all probably heard the Chinese proverb that it’s not the destination that is important but the journey. Well the same can be said of scientific research: it’s not only the results that matter but the methodology and processes that lead us to them.
OK, so this may be a little facetious but the concept is not and it’s the reason why guidelines for the reporting of scientific experiments have been emerging over the past decade across scientific disciplines – including the Minimum Information for Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines.
PCR (polymerase chain reaction) is a technique widely used in molecular biology to identify and quantify DNA. PCR works by targeted amplification of DNA by several orders of magnitude to enable identification and measurement of specific sequences. Quantitative polymerase chain reaction (qPCR), also called real-time polymerase chain reaction, is a laboratory technique based on the PCR, which is used to amplify and simultaneously quantify a targeted DNA molecule.
In order to encourage increased transparency in reported data, the MIQE guidelines were developed and published in 2009. But why is this so important and, five years later, what impact have the guidelines had?
The peer review process for publishing scientific studies relies on the ability of scientists to review the work of others. If they don’t have all of the facts as to how an experiment was carried out, they cannot adequately evaluate the technical standard or results and determine whether a result may be biased or whether differences are due to technical variation. It also means that it is difficult for another scientist to repeat the study or to further develop the research, ultimately hindering the impact that work may have on, for example, understanding the pathology of a given disease.
A team of LGC scientists collaborated on a study evaluating the quality of scientific publications concerned with the measurement of RNA and DNA by qPCR following the introduction of the MIQE guidelines to see if reporting standards had improved. They found that although there have been some advances, even those papers that cite the MIQE guidelines do not always contain all essential technical information.
With reporting guidelines so important to the evaluation of research and the development of further studies, LGC scientists also collaborated on guidance aimed at improving the quality of molecular measurements and led the development of guidelines on the publication of quantitative digital PCR (dPCR) experiments.
dPCR is an extension of conventional PCR methods that allows for very precise quantification of DNA and absolute, rather than the conventional relative, quantification. dPCR has greater sensitivity for differentiating small variations in DNA levels between samples. This has applications in clinical diagnostics for the identification and quantification of rare genetic mutations. dPCR also has the potential to be more reproducible and less susceptible to experimental interferences than current quantitative real-time PCR (qPCR) techniques.
The applications of PCR are widespread including, for example, diagnosis of pathogens, facilitating DNA sequencing for identification of genetic disorders or the detection of genetically modified food products and the implications of failing to adhere to reporting guidelines are vast.
Not only could research resources be wasted at a time when they are increasingly scarce but it also has implications for drug development and disease monitoring.
With so much at stake and with the credibility of their research on the line, why do so many scientists not follow these simple guidelines?
To find out more read “Standardising measurement in molecular biology” on page 4 of the latest issue of Catalyst newsletter.