Assessment of a new qPCR tool for the detection and identification of the root-knot nematode Meloidogyne enterolobii by an international test performance study

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dc.authorid0000-0002-0817-5425
dc.authorid0000-0002-0817-5425
dc.contributor.authorBraun-Kiewnick, Andrea
dc.contributor.authorViaene, Nicole
dc.contributor.authorFolcher, Laurent
dc.contributor.authorOllivier, Fabrice
dc.contributor.authorAnthoine, Geraldine
dc.contributor.authorNiere, Bjoern
dc.contributor.authorKiewnick, Sebastian
dc.date.accessioned2019-08-01T13:38:39Z
dc.date.available2019-08-01T13:38:39Z
dc.date.issued2016
dc.departmentNiğde ÖHÜ
dc.description.abstractRapid and reliable tools for detection and identification of plant parasitic nematodes are needed to prevent the introduction and spread of quarantine nematodes. A fast and simple DNA extraction method for target nematodes in nematode suspensions obtained from soil samples and a new quantitative real-time PCR method (qPCR) for the specific detection, identification and potential quantification of M. enterolobii were tested in an inter-laboratory comparison (ring test) to allow for a thorough evaluation of these molecular diagnostic tools. A test performance study involving seven laboratories was conducted to validate the developed protocols and to identify possible difficulties when implemented by diagnostic laboratories or national reference centers. Validation included test performance in terms of accuracy, analytical specificity, analytical sensitivity, repeatability, and reproducibility as defined by European Plant Protection Organization (EPPO) standard PM7/98. All positive and negative results for detection, identification and specificity were consistent between different laboratories despite different equipment used. Accuracy of real-time PCR was 100 % because test results and accepted reference values were in agreement. Analytical sensitivity results also matched between laboratories independent of the equipment used. The smallest amount of target DNA tested, two second-stage juveniles of M. enterolobii in a background of 500 non-target nematodes, was reliably detected by all labs. In addition, the repeatability and reproducibility of test results between laboratories was 100 %, even at the limit of detection. Thus, the inter-laboratory comparison showed the robustness of the developed methods and confirmed the in-house validation data.
dc.description.sponsorshipEUPHRESCO II framework, project "Development and validation of innovative diagnostic tools for detection and identification of the quarantine nematode Meloidogyne enterolobii in support of integrated plant protection strategies in the EU member states"; Swiss Federal Office of Agriculture; Belgian Federal Agency for the Safety of the Food Chain
dc.description.sponsorshipThis research project was performed within the EUPHRESCO II framework, project "Development and validation of innovative diagnostic tools for detection and identification of the quarantine nematode Meloidogyne enterolobii in support of integrated plant protection strategies in the EU member states",, via a non-competitive funding mechanism. We thank all colleagues for sharing nematode populations and the Swiss Federal Office of Agriculture as well as the Belgian Federal Agency for the Safety of the Food Chain for financial support.
dc.identifier.doi10.1007/s10658-015-0754-0
dc.identifier.endpage108
dc.identifier.issn0929-1873
dc.identifier.issn1573-8469
dc.identifier.issue1
dc.identifier.scopus2-s2.0-84951569063
dc.identifier.scopusqualityQ1
dc.identifier.startpage97
dc.identifier.urihttps://dx.doi.org/10.1007/s10658-015-0754-0
dc.identifier.urihttps://hdl.handle.net/11480/3816
dc.identifier.volume144
dc.identifier.wosWOS:000366635400008
dc.identifier.wosqualityQ2
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.institutionauthor[0-Belirlenecek]
dc.language.isoen
dc.publisherSPRINGER
dc.relation.ispartofEUROPEAN JOURNAL OF PLANT PATHOLOGY
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.subjectReal-time PCR
dc.subjectPerformance assessment
dc.subjectMeloidogyne enterolobii
dc.subjectSensitivity
dc.subjectSpecificity
dc.titleAssessment of a new qPCR tool for the detection and identification of the root-knot nematode Meloidogyne enterolobii by an international test performance study
dc.typeArticle

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