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2017-06-05


Metrology for Quantitative Molecular Biology: Accurate and Reproducible Nucleic Acid Measurements

Mojca Milavec1

1National institute of biology, Department of biotechnology and systems biology, Ljubljana, Slovenia.


Metrology is the science of measurement, embracing both experimental and theoretical determinations at any level of uncertainty in any field of science and technology [1]. It establishes a common understanding of units of measurement and has wide-ranging impacts on a number of sectors, including health, environment, industry, energy, economics, and consumer confidence. While in many fields, like mass and related quantities, length, electricity and magnetism, metrology has long tradition, in other fields, like chemistry and biology metrology is rapidly evolving and expanding along with emerging measurement needs.
 
One of fast developing sub-fields of metrology in biology are nucleic acids (NA) measurements. Accurate, reliable and traceable NA measurements are crucial for research and development in health, biotechnology, food safety, pharmacy, environmental monitoring, and others [2]. In NA measurements, rapid technological advances have occurred over the last few decades. Molecular approaches, such as quantitative polymerase chain reaction (qPCR), digital PCR (dPCR) and sequence analysis, offer the potential for accurate, sensitive, repeatable and reliable detection and/or quantification, however, metrological support is lacking, with issues concerning reproducibility, comparability and traceability of measurements.
 
Many efforts have been made to define measurement procedures and assess key NA measurement parameters. However, challenges remain including the absence of certified reference materials (CRMs) and reference methods, and sometimes undefined validation parameters and unclear use of units in reporting results in certain fields. Moreover, reference/standard methods, key comparison studies, and proficiency testing schemes are not always available [2].
 
At NIB, intensive research has been carried out over the last two decades on the development and assessment of NA measurements in basic research and their application in different fields, in particular for genetically modified organisms (GMOs) and plant pathogens. Based on research and knowledge gained, NIB has become a holder of a national standard (or etalon) in the field of metrology in chemistry/ amount of substance in 2009. Metrological approaches are integrated into the research and development work at NIB. We have observed that, depending on the aim within a particular field, or even of a particular analysis, the requirements of the method performance can vary considerably [2].
 
At the international level NIB is involved in specialised research to support NA measurements in international projects [3] and inter-laboratory comparisons [4] as well as in strategic planning for global comparability of NA measurements [5].
 

  1. www.bipm.org/en/worldwide-metrology
  2. Camloh M, et al. (2015) 17th International Congress of Metrology, 06005
  3. Pavšič J, et al. (2017) Anal Bioanal Chem 409(10):pp. 2601–14
  4. Yoo H et al.(2016) Anal. Chem 88(24), pp.12169–76
  5. www.bipm.org/en/committees/cc/wg/nawg.html


 

 

2006 - University of Ljubljana, Faculty of Medicine, Center for Functional Genomics and Bio-chips.