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Advancing DNA research safely and securely

27 May 2009

Dr Jeremy GarsonDr Jim Huggett

Dr Jeremy Garson (UCL Centre for Virology) and Dr Jim Huggett (UCL Centre for Infectious Diseases and International Health) have been at the heart of developing a new set of guidelines on the way scientists the world over use qPCR – a technology crucial to forensic analysis and diagnosing diseases. Below Dr Huggett explains how and why they went about it.

What is qPCR?

qPCR stands for real-time quantitative polymerase chain reaction. It enables scientists to measure accurately the amount of any particular DNA sequence in a biological sample. The PCR method (in its many guises) is arguably one of the most fundamental technologies for biological and medical research available today.

What is this method used for?

qPCR is used in most aspects of biological, agricultural and medical research where DNA is measured. It’s essential for many aspects of forensic analysis and diagnostic testing, for example to identify if a person has a cancer or an infectious disease).

Why were guidelines needed?

While qPCR is relatively easy to perform, it is very difficult to perform well and in a way that can be repeated exactly by different laboratories.  Consensus guidelines exist for several other laboratory methods, but have not previously been written for qPCR.

What do you hope to achieve with the guidelines?

By developing the MIQE guidelines (Minimum Information for Publication of Quantitative Real-Time PCR Experiments), we aim to enable researchers to perform high-quality qPCR that allows their experiments to be easily understood and repeated by workers in laboratories anywhere in the world. For science to advance swiftly and securely it is essential that the results of experiments can be independently reproduced.

Real-time PCR machine

How did you go about developing the guidelines?

The guidelines have been developed by 12 international experts in qPCR working in academia and industry from several European countries and the United States. The writing of the guidelines was led by Professor Stephen Bustin of Barts and the London School of Medicine and Dentistry. It was stimulated by the recognition that scientific research was being hindered by poorly conducted qPCR experimentation.

UCL’s Dr Jeremy Garson, a leading molecular virologist, and I (having written extensively on molecular measurement), were key in the development of the guidelines by discussing with the co-authors what were felt to be the main failings that needed to be addressed to counter this serious problem. Consensus was eventually reached and the guidelines have now been published in the prestigious international journal, Clinical Chemistry.

What do the guidelines recommend, in lay terms?

The guidelines are a set of essential experimental conditions and technical factors that need to be described by any scientific publication using qPCR. Description of these factors enables the reader of the scientific paper to better judge the validity of the results generated and the conclusions reached.

Who will check that the guidelines are observed?

The guidelines are now available for authors, reviewers and editors to assist the publication process. They aim to enhance transparency by increasing the amount of information available about a study, but are not intended to be excessively rigid or draconian rules on how qPCR must be performed. We have informed editors of key journals of these guidelines and their use is voluntary at this stage, but will considerably strengthen any publication adhering to them.

Images: Dr Jeremy Garson (top); Dr Jim Huggett; and a real-time PCR machine in action