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PCR revisited: a case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar. (Articolo in rivista)

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PCR revisited: a case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar. (Articolo in rivista) (literal)

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On S.L.W.1, Brandt S.M. 1, Cornelius A.J. 1, Fusco V. 2, Quero G.M. 2, Ma?kiw E. 3, 4, Houf K. 5, Bilbao A. 6, Díaz A.I. 6, Benejat L. 7, Megraud F. 7, Collins-Emerson J. 8, French N.P. 8, Gotcheva V. 9, Angelov, A. 9, Alakomi, H.-L. 10, Saarela M. 10, Paulin S.M.* 1 (2013)
PCR revisited: a case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar.
in Quality assurance and safety of crops & foods (Print)
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On S.L.W.1, Brandt S.M. 1, Cornelius A.J. 1, Fusco V. 2, Quero G.M. 2, Ma?kiw E. 3, 4, Houf K. 5, Bilbao A. 6, Díaz A.I. 6, Benejat L. 7, Megraud F. 7, Collins-Emerson J. 8, French N.P. 8, Gotcheva V. 9, Angelov, A. 9, Alakomi, H.-L. 10, Saarela M. 10, Paulin S.M.* 1 (literal)

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1Christchurch Science Centre, Institute of Environmental Science and Research (ESR), Food Programme, 27 Creyke Road, Ilam, 8041, Christchurch, New Zealand 2National Research Council of Italy, Institute of Sciences and Food Protection (CNR-ISPA), Via Amendola 122/o, 70126 Bari, Italy 3National Food and Nutrition Institute (NFNI), Powsi?ska 61/63, 02-093 Warsaw, Poland 4Department of Food and Consumer Articles Research, National Institute of Public Health - National Institute of Hygiene, ul. Chocimska 24, 00-791 Warszawa, Poland 5Department of Veterinary Public Health and Food Safety, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium 6Gaiker-IK 4 Zentru Teknologikoa, Teknologi Parkea, 202 Eraikina, 48170 Zamudio, Bizkaia, Spain 7Laboratoire de Bacteriologie (INSERM U853), Campylobacter National Reference Centre, University Bordeaux Segalen, 146 Rue Leo Saigent, 33076 Bordeaux, France 8mEpiLab, Hopkirk Research Institute, Massey University, IVABS, Tennent Drive, 4442 Massey, New Zealand 9Department of Biotechnology, University of Food Technologies, 26 Maritza Blvd, 4002 Plovdiv, Bulgaria 10VTT, Technical Research Centre of Finland, Tietotiez, 02044 Espoo, Finland (literal)

Titolo

PCR revisited: a case for revalidation of PCR assays for microorganisms using identification of Campylobacter species as an exemplar. (literal)

Abstract

We re-examined the sensitivity and specificity of 31 PCR assays (including four commercially available and developed in-house methods) for the identification of Campylobacter species, with particular reference to taxa described since 2004, which are closely related to C. jejuni and C. coli, the pathogenic species of most interest. Each of the assays was used by at least one of the participating nine laboratories in eight countries. The sensitivity and specificity of these PCR assays examined varied considerably and ranged from 100% to 0% for sensitivity and 100% to 55% for specificity. None of the three assays examined for C. lari were successful in detecting all strains of this species, possibly reflecting its complex taxonomy. A number of assays for C. jejuni, C. coli, and a subgroup of enteropathogenic campylobacters, were found to yield false positive results for Campylobacter species described since PCR tests were reported, including C. cuniculorum, C. subantarcticus, C. peloridis and C. volucris. Our study supports the need for attention to detail in initial PCR assay design and evaluation, and also for on-going revalidation of laboratory assays to ensure that diagnoses are correct. Recommendations to guide the revalidation process are presented. (literal)

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