Development and evaluation of two TaqMan assays for generic detection of Dickeya species

Zijlstra, C.; Haas, Lia Groenenboom – De; Krijger, M.C.; Verstappen, E.C.P.; Warris, Sven; Haan, Eisse de; Wolf, Jan van der

doi:10.1007/s10658-019-01876-y

Cited by 6 publications

(2 citation statements)

References 14 publications

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“…Existing literature reports two TaqMan qPCR assays for general Dickeya species detection, using D. solani bacterial DNA in plant samples, with a sensitivity of 100 fg/reaction (Zijlstra et al, 2020). In our newly developed primer/probe pairs DsowF1/wR1/Dso-P1 and DsowF2/wR2/Dso-P2 detecting as low as 10 fg (~1.8 copies).…”

Section: Discussionmentioning

confidence: 99%

Development and Validation of Genome-Informed and Multigene-based qPCR and LAMP Assays for Accurate Detection ofDickeya solani: A Critical Quarantine Pathogen Threatening Potato Industry

Dobhal,

Santillana,

Stulberg

et al. 2024

Preprint

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Dickeya solani, one of the most aggressive pectinolytic phytopathogens, causes blackleg disease in potatoes, resulting in significant economic losses and adversely impacting one of the world's most important food crops. The diagnostics methods are critical in monitoring the latent infection for international trade of potato seed-tubers and in implementation of control strategies. Our study employed a whole-genome comparative approach, identifying unique target gene loci (LysR and TetR family of transcriptional regulators gene regions), design loop-mediated isothermal amplification (LAMP) and a multi-gene-based multiplex TaqMan qPCR assays for specific detection and differentiation of D. solani. Both methods underwent meticulous validation with extensive inclusivity and exclusivity panels, exhibiting 100% accuracy and no false positives or negatives. The LAMP method demonstrated the detection limit of 100 fg and 1 CFU per reaction using pure genomic DNA and crude bacterial cell lysate, respectively. The qPCR detection limit was 1 pg, 100 fg and 10 fg with Quadplex, triplex and singleplex, respectively. None of the assay showed any inhibitory effect after adding host DNA (in qPCR) or crude lysate (in LAMP). The assays proved robust and reproducible in detecting the target pathogen in infected samples, with the LAMP assay being field-deployable due to its simplicity and rapid results acquisition within approximately 8.92 minutes. The reproducibility was confirmed by performing the assay in two independent laboratories. These developed assays offer a robust, rapid, and reliable solution for routine testing, with applications in phytosanitary inspection and epidemiological studies.

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Section: Discussionmentioning

confidence: 99%

Development and Validation of Genome-Informed and Multigene-based qPCR and LAMP Assays for Accurate Detection ofDickeya solani: A Critical Quarantine Pathogen Threatening Potato Industry

Dobhal,

Santillana,

Stulberg

et al. 2024

Preprint

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“…These include TaqMan assays for the detection and quantification of all SRP, Dickeya spp. (Zijlstra et al 2019)…”

Section: Real-time (Quantitative) Pcrmentioning

confidence: 99%

Isolation, Detection and Characterization of Pectobacterium and Dickeya Species

Wolf

Cahill

Gijsegem

et al. 2021

Plant Diseases Caused by Dickeya and Pectobacterium Species

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Article 25fa states that the author of a short scientific work funded either wholly or partially by Dutch public funds is entitled to make that work publicly available for no consideration following a reasonable period of time after the work was first published, provided that clear reference is made to the source of the first publication of the work.This publication is distributed under The Association of Universities in the Netherlands (VSNU) 'Article 25fa implementation' project. In this project research outputs of researchers employed by Dutch Universities that comply with the legal requirements of Article 25fa of the Dutch Copyright Act are distributed online and free of cost or other barriers in institutional repositories. Research outputs are distributed six months after their first online publication in the original published version and with proper attribution to the source of the original publication.

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Dickeya solani (black leg disease of potato)

Osdaghi¹

2022

CABI Compendium

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Development and evaluation of two TaqMan assays for generic detection of Dickeya species

Cited by 6 publications

References 14 publications

Development and Validation of Genome-Informed and Multigene-based qPCR and LAMP Assays for Accurate Detection ofDickeya solani: A Critical Quarantine Pathogen Threatening Potato Industry

Development and Validation of Genome-Informed and Multigene-based qPCR and LAMP Assays for Accurate Detection ofDickeya solani: A Critical Quarantine Pathogen Threatening Potato Industry

Isolation, Detection and Characterization of Pectobacterium and Dickeya Species

Dickeya solani (black leg disease of potato)

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