WG Leader: Dario de Medici, ISS, Italy

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Members: Jana Hajslova, VSCHT, Czech Republic; Angel Angelov, Velitchka Gotcheva, UFT, Bulgaria; Martino Barbanera, COOP, Italy; Andreas Höhl, BOKU, Austria; Mindaugas Malakauskas, LVA, Lithuania; Peter Siekel, VUP, Slovakia; To Kim Anh, HUST, Vietnam; Estella Prukner-Radovcic, VEF, Croatia; Rickard Knutsson, SVA, Sweden.

Research topics for the future:

Standardization and validation of molecular method protocols: Standardization of modern technologies such as molecular methods can be achieved either by the development of specific standards for novel technologies or by elaborating standards laying out the general guidelines for applying PCR in food analysis.

Developing new rapid technologies for emerging microbiological risks: New technologies and methods also have to reflect the emerging needs for added information, such as sub-species identification, presence of genes important for virulence or antibiotic resistance, but also to detect important food-borne pathogens for which no classical counterparts are available, namely food-borne pathogenic viruses.

Workshops organised

WG9 also organised the EU-Vietnam Workshop that took place in Hanoi in March 2014. See FOODSEG Events.

Relevant publications by the Group:

Title (and if available Link): A decade with nucleic acid-based microbiological methods in safety control of foods.

Link: http://onlinelibrary.wiley.com/doi/10.1111/lam.12283/abstract;jsessionid=64D7112030CF4605818C29DED40EAC88.f03t01; Authors: Kuchta T1, Knutsson R, Fiore A, Kudirkiene E, Höhl A, Horvatek Tomic D, Gotcheva V, Pöpping B, Scaramagli S, To Kim A, Wagner M, De Medici D. ; Hourna Lett Appl Microbiol. 2014 May 12. doi: 10.1111/lam.12283. [Epub ahead of print]

Abstract: In the last decade, nucleic acid-based methods gradually started to replace or complement the culture-based methods and immunochemical assays in routine laboratories involved in food control. In particular, real-time polymerase chain reaction (PCR) was technically developed to the stage of good speed, sensitivity and reproducibility, at minimized risk of carry-over contamination. Basic advantages provided by nucleic acid-based methods are higher speed and added information, such as subspecies identification, information on the presence of genes important for virulence or antibiotic resistance. Nucleic acid-based methods are attractive also to detect important foodborne pathogens for which no classical counterparts are available, namely foodborne pathogenic viruses. This review briefly summarizes currently available or developing molecular technologies that may be candidates for involvement in microbiological molecular methods in the next decade. Potential of nonamplification as well as amplification methods is discussed, including fluorescent in situ hybridization, alternative PCR chemistries, alternative amplification technologies, digital PCR and nanotechnologies.

Title (and if available Link): ) Rapid methods for quality assurance of foods: the next decade with Polymerase Chain Reaction (PCR) based food monitoring

Author: D. De Medici, T. Kuchta, R. Knutsson, A. Angelov, B. Auricchio, M. Barbanera, C. Diaz-Amigo, A. Fiore, E. Kudirkiene, A. Hohl, D. Horvatek Tomic, V. Gotcheva, B. Popping, E. Prukner-Radovcic, S. Scaramaglia, P. Siekel, K.A. To, M. Wagner. Journal Food Analytical Methods accepted for the publication.

Abstract: Microbiological analysis is an integral part of food quality control, as well as of the management of food chain safety. Microbiological testing of foodstuffs complements the preventive approach to food safety activities based mainly on implementation and application of the concept of Hazard Analysis and Critical Control Points (HACCP). Traditional microbiological methods are powerful, but lengthy and cumbersome and therefore not fully compatible with current requirements. Even more, pathogens exist that are fastidious to cultivate or uncultivable at all. Besides immunological tests, molecular methods, specifically those based on polymerase chain reaction (PCR), are available options to meet industry and enforcement needs. The clear advantage of PCR over all other rapid methods is the striking analytical principle that is based on amplification of DNA, a molecule being present in every cell prone to multiply. Just by changing primers and probes, different genomes such as bacteria, viruses or parasites can be detected. A second advantage is the ability to both detect and quantify a biotic contaminant. Some previously identified obstacles of implementation of molecular methods have already been overcome. Technical measures became available that improved robustness of molecular methods, and equipment and biochemicals became much more affordable. Unfortunately, molecular methods suffer from certain drawbacks that hamper their full integration to food safety control. Those encompass a suitable sample pre-treatment especially for a quantitative extraction of bacteria and viruses from solid foods, limited availability of appropriate controls to evaluate the effectiveness of the analytical procedure; the current inability of molecular methods to distinguish DNA from viable cells and DNA from dead or non-cultivable cells, and the slow progress of international harmonization and standardization, that limit full acceptance of PCR-based methods in food control. The aim of this review is to describe the context and the prospects of PCR-based methods, as well as trends in research and development aimed at solving the next decade challenges in order to achieve full integration of molecular methods in food safety control.