Premium
Euphresco inter‐laboratory comparison (2009–2012) on detection of Clavibacter michiganensis subsp. sepedonicus and Ralstonia solanacearum in potato tubers: proposal to include TaqMan ® real‐time PCR as a primary (core) screening test in EU / EPPO standard methods
Author(s) -
Vaerenbergh J.,
Müller P.,
Elphinstone J. G.,
Vreeburg R. A. M.,
Janse J. D.
Publication year - 2017
Publication title -
eppo bulletin
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.327
H-Index - 36
eISSN - 1365-2338
pISSN - 0250-8052
DOI - 10.1111/epp.12352
Subject(s) - clavibacter michiganensis , ralstonia solanacearum , taqman , bacterial wilt , biology , european commission , european union , real time polymerase chain reaction , microbiology and biotechnology , horticulture , pathogen , genetics , gene , business , economic policy
In the European Union ( EU) potato production is surveyed for Clavibacter michiganensis subsp. sepedonicus (potato ring rot) and Ralstonia solanacearum (potato brown rot) under Commission Directives 93/85/EEC with its amendment 2006/56/ EC and 98/57/EEC with its amendment 2006/63/ EC . A regular update of the Directives is required in view of developments in understanding of the biology of these organisms and the diagnostics recommended for their detection and identification. Three inter‐laboratory tests ( ILT 1, ILT 2 and ILT 3) were performed from 2009 to 2012 as part of a Euphresco Phytosanitary ERA ‐ NET project to assess performance of current official methods for C. michiganensis subsp. sepedonicus and R. solanacearum . A major aim of the ILT s was to generate data on the performance of real‐time PCR protocols to support their introduction as primary (core) screening tests for both pathogens. In ILT 1, 29 laboratories from 23 countries participated, in ILT 2, 23 laboratories from 18 countries and in ILT 3 42 laboratories from 24 countries. Relative accuracies for real‐time PCR tests averaged 92% for R. solanacearum and 96% for C. michiganensis subsp. sepedonicus ) and compared with existing primary (core) screening tests (immunofluorescence, conventional PCR , semi‐selective plating and bioassay) in terms of analytical sensitivity, analytical specificity and robustness. It was concluded that all methods tested, including real‐time PCR , can be considered as equivalent. Therefore TaqMan ® real‐time PCR is recommended for inclusion in EU Directives and EPPO Standards as a reliable primary (core) screening method.