Euphresco Sendo: An international laboratory comparison study of molecular tests for Synchytrium endobioticum detection and identification

Vossenberg, Bart van de; Westenberg, Marcel; Adams, Ian; Afanasenko, Olga; Besheva, Ani; Boerma, M.; Choiseul, J.W.; Dekker, Toos; Flath, Kerstin; Gent-Pelzer, Marga van; Heungens, Kurt; Karelov, A. V.; Kibildiene, Ilona; Przetakiewicz, Jarosław; Schlenzig, A.; Yakovleva, V. A.; Leeuwen, Gerard van

doi:10.1007/s10658-017-1411-6

Cited by 10 publications

(6 citation statements)

References 16 publications

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“…One of those, the ITS2 test described by Van Gent‐Pelzer et al (2010), was part of the Euphresco TPS. In combination with the DNeasy Plant Mini kit, this real‐time PCR was not fit for the detection of spores in soil (Van de Vossenberg et al, 2018). With the introduction of the filtration set‐up to concentrate the spores from the saturated CaCl 2 solution and the DNeasy PowerSoil kit with the bead‐beater (this study), an increased sensitivity was obtained for the ITS2 real‐time PCR, i.e.…”

Section: Discussionmentioning

confidence: 99%

“…As part of the Euphresco Sendo project, an interlaboratory test performance study (TPS) was performed to compare several molecular tests for the detection of S. endobioticum spores in warted tubers and soil suspensions (Van de Vossenberg et al, 2018). The tests included were an S. endobioticum-specific conventional PCR (Van den Boogert et al, 2005), an S. endobioticumspecific real-time PCR (van Gent-Pelzer et al, 2010), and a pathotype 1(D1) specific real-time PCR (Bonants et al, 2015).…”

Section: Bac Kgrou N Dmentioning

confidence: 99%

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Comparison of real‐time PCR tests for the detection of Synchytrium endobioticum resting spores in soil and their potential application in descheduling of previously infested plots

Vossenberg¹,

Braak²,

Warbroek³

et al. 2021

EPPO Bulletin

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Plots infested with Synchytrium endobioticum, the causal agent of potato wart disease, are scheduled, resulting in prohibition of potato cultivation of and cultivation of plants for planting. On account of robust resting spores that are present in infested soils, plots remain scheduled for 20 years. After this period, plots are intensively sampled and the presence of resting spores in soil is determined by direct examination. However, this method is very time-consuming and labour-intensive.In this paper, validation data for the molecular detection of resting spores in soil and its use to screen soils before direct examination are reported. In addition to the samples in the validation study, over 670 routine diagnostic samples were analysed by direct examination and molecular tests in parallel. Using an improved methodology, increased sensitivity was obtained relative to results reported from an interlaboratory comparison study in 2018, namely 7 spores per sample instead of 500 spores per sample. Molecular screening of the soil samples and direct examination of those samples testing positive was estimated to reduce the total hands-on time by half when compared to direct examination of all samples. We recommend inclusion of molecular detection in the update of EPPO PM 3/59 (3) and PM 7/28(2), and suggest that when all subsamples of a plot are negative, no additional direct examination or bio-assays are needed for descheduling (i.e. for releasing previously infested plots from official control). Comparaison de tests PCR en temps réel pour la détection de spores dormantes de Synchytrium endobioticum dans le sol et leur application potentielle pour la déréglementation des parcelles précédemment infestésLes parcelles infestés par Synchytrium endobioticum, l'agent causal de la galle

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Section: Discussionmentioning

confidence: 99%

Section: Bac Kgrou N Dmentioning

confidence: 99%

Comparison of real‐time PCR tests for the detection of Synchytrium endobioticum resting spores in soil and their potential application in descheduling of previously infested plots

Vossenberg¹,

Braak²,

Warbroek³

et al. 2021

EPPO Bulletin

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“…Bioassays using a set of differential potato varieties have been developed for the identification of the most common pathotypes present in Europe (EU and non-EU countries), i.e. 1(D1), 2 (G1), 6(O1) and 18(T1) and molecular methods are available for distinguishing pathotype 1(D1) from the other three most common pathotypes present in Europe (van de Vossenberg et al, 2017;EPPO, online).…”

Section: Detection and Identification Of The Pestmentioning

confidence: 99%

Pest categorisation of Synchytrium endobioticum

Jeger¹,

Bragard²,

Caffier³

et al. 2018

EFS2

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The Panel on Plant Health performed a pest categorisation of the fungus Synchytrium endobioticum, the causal agent of potato wart disease, for the European Union (EU). The identity of the pest is well established and reliable methods exist for its detection and identification. S. endobioticum is present in most continents. The pest is listed in Annex IAII of Directive 2000/29/EC and is present with a restricted and fragmentary distribution in the EU. The major host is Solanum tuberosum (potato), but in Mexico, the pest also affects wild Solanum spp. S. endobioticum could potentially enter the EU through multiple pathways associated with soil as substrate for non‐host plants, contaminant or commodity. The presence of the pest in 16 EU Member States characterised by different climatic conditions suggests that it could establish in the rest of the EU. The disease induces the formation of warts on potato tubers, stolons and stem bases reducing plant growth and yield and making tubers unmarketable. Additional losses may occur during storage. The only available strategy to control the disease and prevent it from spreading is the application of strict phytosanitary measures and the cultivation of potato varieties resistant to the pathotype(s) present in the infested field(s). Specific phytosanitary measures exist (Council Directive 69/464/EEC) for the control of potato wart disease in the EU. The main uncertainties refer to the distribution and host range of the pest, and the importance of some pathways of entry. S. endobioticum meets all the criteria assessed by EFSA for consideration as potential Union quarantine pest. The criteria for considering S. endobioticum as a potential Union regulated non‐quarantine pest are not met since, in addition to potato seed tubers, soil (as commodity, substrate or contaminant) and ware potato tubers are major means of spread.

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“…Potato wart is included in the list of quarantine diseases in 53 countries worldwide (EPPO Global Database, GB, 2021). At present, there have been 39 pathotypes of S. endobioticum described (Przetakiewicz J, 2015b;Przetakiewicz J, 2017;van de Vossenberg B et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

Efficiency of detecting resting spores of Synchytrium endobioticum (potato wart disease) in infested sites of Ukraine and Georgia

Zelya¹,

Janse²,

Janse³

et al. 2021

Agric. sci. pract.

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Aim. To determine the occurrence and persistence of Synchytrium endobioticum, resting spore contamination in a small survey of (known infested) potato plots in Ukraine and Georgia; to compare the detection efficiency for resting spores (winter sporangia) of S. endobioticum using an extraction method, routinely applied in Ukraine, based on the use of sodium iodide (NaI) and an extraction method largely based on EPPO Standard PM 7/28(2) (2017), using kaolin and calcium chloride (CaCl2) for extraction. Methods. The examination of fields, aimed at detecting of S. endobioticum in 22 infested plots in Georgia, was conducted following the standard European Plant Protection Organisation (EPPO) phytosanitary procedure PM 3/59(3) (2017), and in Ukraine in 11 plots according to the Ukrainian Standard ‘Methodological recommendations on sampling during quarantine inspection and evaluation’ (Omeluta V P et al, 1996). Resting spores were extracted using kaolin and CaCl2 (following largely EPPO Standard PM 7/28(2) 2017), and floatation in a solution of NaI (Zelya et al, 2005), respectively. The content of soil organic matter (SOM), collected in potato wart infested plots in Ukraine and Georgia, was determined according to the ‘Method of laboratory determination of the content of organic matter’, largely following the method of Tyurin to determine total soil organic carbon (Jankauskas B et al, 2006). The persistence or decline of potato wart in infested plots was evaluated, based on our own observations and the data of the national phytosanitary services (https://dpss.gov.ua; http://agr.georgia.gov). The results of the study were subjected to statistical analysis, using Statistica 5 software. Results. It was found that under a low up to high level of organic matter in soil samples (2.0–2.7 and 3.1–3.9 %, respectively) and a moderate to high level of inoculum in soil (3–15 up to 41–65 resting spores/g soil, respectively) there were no significant differences in the efficiency of two extraction methods under consideration: the floatation in a solution of NaI and the application of kaolin and CaCl2. However, at a low number of resting spores present in soil (1–2 resting spores/g soil), the efficiency of the method using NaI decreased statistically significant by 20–30 % as opposed to the method using kaolin/CaCl2. A relatively high level of soil contamination with resting spores was found in 11 investigated plots of 4 Ukrainian regions: 41–46 resting spores/g soil in the Ivano-Frankivsk Region, 49 in the Lviv Region, 40–65 in the Zakarpattia and 52–65 Chernivtsi Regions. The majority of the 22 investigated Georgian plots showed a low inoculum level (1–7 resting spores/g soil) and only in one village their level amounted to 15 resting spores/g soil (Uchguli village, Mestia municipality). Conclusions. The method applying sodium iodide was found to be comparable to the method applying kaolin and CaCl2 under conditions of moderate to high inoculum levels (15–65 resting spores/g soil) but the latter method was more efficient under conditions of high content of organic matter and very low inoculum level (

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Euphresco Sendo: An international laboratory comparison study of molecular tests for Synchytrium endobioticum detection and identification

Abstract: An international test performance study (TPS) was organised to generate validation data for three molecular Synchytrium endobioticum tests: van den Boogert et al.

Cited by 10 publications

References 16 publications

Comparison of real‐time PCR tests for the detection of Synchytrium endobioticum resting spores in soil and their potential application in descheduling of previously infested plots

Comparison of real‐time PCR tests for the detection of Synchytrium endobioticum resting spores in soil and their potential application in descheduling of previously infested plots

Pest categorisation of Synchytrium endobioticum

Efficiency of detecting resting spores of Synchytrium endobioticum (potato wart disease) in infested sites of Ukraine and Georgia

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