Certification of a reference material of Campylobacter coli and jejuni (CNET068 and CNET112) agarose plugs for PFGE: IRMM-313

This report describes the production of IRMM-313, a Campylobacter coli and jenuni gDNA material certified for the size of the DNA fragments obtained by enzymatic restriction and Pulsed Field Gel Electrophoresis (PFGE). The material was produced following ISO Guide 34:2009. The CRM was produced from cultures of Campylobacter coli CNET068 and jejuni CNET112 which were pooled and processed into agarose plugs suited for PFGE. The bacterial cells were lysed as to release the gDNA within the plug. Between unit-homogeneity and stability during dispatch and storage were assessed in accordance with ISO Guide 35:2006. The material was characterised by an intercomparison among laboratories of demonstrated competence and complying to ISO/IEC 17025. Technically invalid results were removed but no outlier was eliminated on statistical grounds only. The certified values were obtained by PFGE. Uncertainties of the certified values were calculated in compliance with the Guide to the Expression of Uncertainty in Measurement (GUM). The material is intended for quality control and assessment of method performance. As any reference material, it can also be used for control charts or validation studies. The CRM is available in plastic screw cap vials containing 1 plug suspended in Tris EDTA buffer solution. The minimum amount of sample recommended to be used is ½ plug.JRC.D.2-Standards for Innovation and sustainable Developmen

Certification of a Reference Material with Enterococcus Faecalis (CIP 106877) at a Target Level of 1000 Colony Forming Units per Material Sphere, IRMM-355

This report describes the certification of a reference material (IRMM-355) of Enterococcus faecalis. Certified Reference Materials (CRMs) for microbiological analysis are indispensible tools for development and validation of detection methods and for the implementation and support of internal and external quality control in the area of microbiological analysis. The content of each vial of IRMM-355 is one material sphere containing Enterococcus faecalis. The homogeneity and stability (at -20 °C and -70 °C) of the batch was assessed by counting colony forming units (cfu) per material sphere on horse blood agar (BA) and Slanetz and Bartley agar (SB agar). The material is stable when stored at a maximum temperature of -20 °C for up to 18 months. The batch was characterised by six laboratories to establish a certified value of cfu per material sphere on BA and SB agar. The certified value for BA is 890 cfu with an expanded uncertainty of 135. The certified value for SB agar is 823 cfu with an expanded uncertainty of 126. In both cases, a coverage factor k = 2 is used, corresponding to a level of confidence of about 95 %. The identity of the bacteria was confirmed by DNA sequence analysis of the coding region for the Enterococcus faecalis endocarditis antigen (efaA).JRC.D.2-Reference material

Certification of a Reference Material Consisting of Genomic DNA Inserts of Bacillus Subtilis DSM 5750 for PFGE, IRMM-312

This report describes the production and certification of IRMM-312, a reference material of genomic DNA (gDNA) of Bacillus subtilis DSM 5750 in agarose inserts. This CRM (IRMM-312) is intended to be used for the taxonomic identification of authorised probiotic feed additives by Pulsed Field Gel Electrophoresis (PFGE). The batch was found to be homogeneous and the material is stable at 4 °C. The batch was characterised by five laboratories determining the PFGE pattern of a SfiI restriction enzyme digest of IRMM-312. The pattern should be considered as a whole and restriction fragments in the size interval from 15 kb to 97 kb are certified for their fragment length.JRC.D.2-Reference material

Certification of a Reference Material with Eschericia Coli O157 (NCTC 12900) at a level of 4 Colony forming Unit per Material Sphere, IRMM-351

This report describes the certification of a reference material (IRMM-351) of Escherichia coli O157. Certified Reference Materials (CRMs) for microbiological analysis are indispensable tools for development and validation of detection methods and for the implementation and support of internal and external quality control in the area of microbiological analysis. Each vial contains one material sphere of E. coli O157. The homogeneity and stability (at 4 °C, -20 °C and -70 °C) of the batch was assessed by monitoring colony forming units (cfu) on nutrient agr (NA) and eterohemolysin agar (EhlyA) of selected vials by colony counting. The material is not stable at 4 °C but no instability was detected when stored at -20 °C for up to 12 months and at -70 °C for up to 54 months. The batch was characterised by six laboratories to determine a certified value of cfu per vial on NA and EhlyA. The certified value is 4 cfu on both agars with an expanded uncertainty of 2 using a coverage factor k = 2, corresponding to a level of confidence of about 95 %. DNA sequence analysis of the coding region for the fliC gene identified the material as E. coli O157.JRC.D.2-Reference material

Certification of a Reference Material with Salmonella Enteritidis (NCTC 12694) at a Level of 5 Colony Forming Units on Nutrient Agar and 4 Colony Forming Units on Xylose Lysine Deoxycholate Agar, IRMM-352

This report describes the certification of a reference material (IRMM-352) of Salmonella Enteritidis. Certified Reference Materials (CRMs) for microbiological analysis are indispensable tools for development and validation of detection methods and for the implementation and support of internal and external quality control in the area of microbiological analysis. Each vial contains one material sphere of S. enteritidis. The homogeneity and stability (at 4 °C, -20 °C and -70 °C) of the batch was assessed by monitoring colony forming units (cfu) on nutrient agar (NA) and xylose lysine deoxycholate (XLD) agar of selected vials by colony counting. The material is not stable at 4 °C byt no instability was detected when stored at -20 °C for up to 12 months and at -70 °C for up to 54 months. The batch was characterised by six laboratories to determine a certified value of cfu per vial on NA and XLD agar. A quite considerable performance difference of both agars was observed. The certified value is 5 cfu on NA with an expanded uncertainty of 2 and 4 cfy on XLD agar with an expanded incertainty of 2 using a coverage factor k = 2, corresponding to a level of confidence of about 95 %. DNA sequence analysis of the coding region for the sefA gene identified the material as S. enteritidis.JRC.D.2-Reference material

Treatment of pain following cancer : applying neuro-immunology in rehabilitation practice

Aim: Pain is the second most frequent persistent symptom following cancer treatment. This article aims at explaining how the implementation of contemporary pain neuroscience can benefit rehabilitation for adults following cancer treatment within an evidence-based perspective. Materials and methods: Narrative review. Results: First, pain education is an effective but underused strategy for treating cancer related pain. Second, our neuro-immunological understanding of how stress can influence pain highlights the importance of integrating stress management into the rehabilitation approach for patients having cancer-related pain. The latter is supported by studies that have examined the effectiveness of various stress management programmes in this population. Third, poor sleep is common and linked to pain in patients following cancer treatment. Sleep deprivation results in a low-grade inflammatory response and consequent increased sensitivity to pain. Cognitive behavioural therapy for sleep difficulties, stress management and exercise therapy improves sleep in patients following cancer treatment. Finally, exercise therapy is effective for decreasing pain in patients following cancer treatment, and may even decrease pain-related side effects of hormone treatments commonly used in cancer survivors. Conclusions: Neuro-immunology has increased our understanding of pain and can benefit conservative pain treatment for adults following cancer treatment

Certification of a Reference Material of Purified Genomic DNA from Campylobacter Jejuni (NCTC 11351), Certified Reference Material IRMM-448

This report describes the production and certification of a reference material of purified genomic DNA (gDNA) of Campylobacter jejuni (NCTC 11351). This CRM (IRMM-448) supports the harmonisation and validation of PCR methods by their use as taxonomic controls in PCR reactions. The homogeneity and stability (at three different temperatures for up to 4 weeks) of the batch was assessed by monitoring the amplification of the ceuE gene by PCR, by agarose gel electrophoresis of gDNA and determination of the mass of gDNA per vial. The material is stable at -20 °C. The authenticity of the gDNA was confirmed by DNA sequence analysis of the ceuE gene. Each vial contains a certified mass of 118 ng freeze-dried gDNA, with an expanded uncertainty (UCRM) of 65 ng (k = 2).JRC.D.2-Reference material

Certification of Reference Materials for Detection of the Human Prothrombin Gene G20210A Sequence Variant

There is a need for reference materials in the field of genetic testing for verification of tests results obtained in patients and probands. New types of certified reference materials (CRMs) for genetic testing of the human prothrombin gene G20210A mutation are available. Homogeneity, stability and fitness for the purpose of the plasmids could be demonstrated and no evidence was found that they would not work with other methods as long as these are targeting the whole or parts of the prothrombin gene fragment inserted into the plasmids. The described CRMs support the efforts of the international community in development, validation and harmonisation of tests for molecular genetic testingJRC.D.2-Reference material

CERTIFICATION REPORT: The Certification of the PFGE fragment sizes of Listeria monocytogenes (strain H2446) DNA in agarose plugs: ERM®-AD624

Summary This report describes the production of ERM®-AD624, a Listeria monocytogenes DNA material certified for the size of the DNA fragments obtained by enzymatic restriction digestion and Pulsed Field Gel Electrophoresis (PFGE). This material was produced following ISO Guide 34:2009 [ ] and is certified in accordance with ISO Guide 35:2006 [ ]. The CRM was produced from a culture of Listeria monocytogenes strain H2446 and processed into agarose plugs suitable for PFGE. The bacteria were lysed to release the DNA within the plugs. Between unit-homogeneity and stability during dispatch and storage were assessed in accordance with ISO Guide 35:2006. The material was characterised by an interlaboratory comparison of laboratories of demonstrated competence and adhering to ISO/IEC 17025. Technically invalid results were removed but no outliers were eliminated on statistical grounds only. Uncertainties of the certified values were calculated in accordance with the Guide to the Expression of Uncertainty in Measurement (GUM). The material is intended for quality control and assessment of method performance. As with any reference material, it can be used for establishing control charts or validation studies. The CRM is available in plastic screw cap vials containing one plug suspended in TE solution. The minimum amount of sample to be used is the whole CRM.JRC.F.6-Reference Material

First Certified Reference Materials for Molecular Fingerprinting of Two Approved Probiotic Bacillus Strains

At the present probiotic bacteria are widely used in human and animal nutrition because they beneficially influence the balance of the intestinal flora of the host. Positive effects related to probiotics are various and include treatment of juman food allergy and other diseases, reduction in use of antibiotics and improved animal growth and feed conversion; To protect human and animal health and to improve consumer confidence, a strict legislation on the use of probiotics exists within the European Union (EU). Official controls by national authorities are performed to ensure verification of compliance with feed and food law. Apart from the risk of using unauthorized strains, mislabelling is a knwon problem, partly because of the use of phenotyping or genotyping methods with a lack of discriminative power. In addition to official controls, private controls by food and feed producing companies are important in th eframe of protection of patented strains and industrial property rights. To support these applications, IRMM has developed certified reference materials (CRMs) consisting of genomic DNA inserts of B. subtilis DSM 5749 and B. licheniformis DSM 5750, two strains that received EU approval. In this study we investigated the use of these CRMs, IRMM-311 and IRMM-312, for the detection and unambiguous discrimination of Bacillus strains by pulsed-field gel electrophoresis (PFGE). Identical fingerprints were obtained for the cRMs and control strains isolated from the feed additive Bioplus 2B R. On the other hand a distinction could be made from other not approved B. licheniformis and B. subtilis strains. The reference materials discussed in this study are the first CRMs base on a whole bacterial genome and suitable for PFGE. They offer perspectives for applications in other domains such as analysis of foodborne pathogens in outbreaks or routine analysis.JRC.D.2-Reference material