Comparative evaluation of lateral flow immunoassays, LAMP, and quantitative PCR for diagnosis of fire blight in apple or
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ORIGINAL ARTICLE
Comparative evaluation of lateral flow immunoassays, LAMP, and quantitative PCR for diagnosis of fire blight in apple orchards Jugpreet Singh 1 & Della Cobb-Smith 1 & Elizabeth Higgins 2 & Awais Khan 1 Received: 11 March 2020 / Accepted: 26 August 2020 # Società Italiana di Patologia Vegetale (S.I.Pa.V.) 2020
Abstract Fire blight remains a serious threat to commercial apple production in the USA and worldwide. Other diseases and spray damage can result in fire blight-like symptoms that can lead to misdiagnosis and affect disease management strategies. Accurate and timely detection of the fire blight pathogen, Erwinia amylovora, is extremely important to deploy appropriate and timely measures to reduce fire blight epidemics in commercial apple orchards. We tested two commercial lateral flow immunoassays (AgriStrip®, and Pocket Diagnostics kit), Loop mediated isothermal amplification (LAMP), and quantitative PCR (qPCR) to diagnose E. amylovora infected samples in lab and field settings. The AgriStrip® and Pocket Diagnostics kits were able to detect actively growing bacteria up to ×106 cfu/ml bacterial concentration. Pocket Diagnostics kit had less specificity and showed positive tests for E. pyrifolia in addition to E. amylovora. The LAMP assay showed high specificity for E. amylovora and was able to detect up to ×103 cfu/ml bacterial concentrations. The qPCR assay was also able to detect bacterial cells up to ×10−3 cfu/ ml bacterial concentration with highly specific E. amylovora detection. Grower surveys and comparative cost-benefit analysis indicated that immunoassay kits are less expensive, easier to use, and require less technical expertise for on-site fire blight diagnosis than LAMP and qPCR. However, the choice of a specific diagnostic assay depends on the time, sensitivity, and specificity required for the detection of fire blight and its management. Keywords Pathogen detection . Pathogen diagnosis . Loop mediated isothermal amplification . Enzyme-linked Immuno sorbent assay . Immunoassay . Erwinia amylovora
Introduction Fire blight-causing bacteria, Erwinia amylovora, poses a great threat for commercial apple production worldwide (van der Zwet et al. 2012; Norelli et al. 2003). Fire blight has caused significant losses in United States alone, with over $42 million in Michigan in 2000 (Longstroth 2001) and $68 million in northern Oregon and Washington Electronic supplementary material The online version of this article (https://doi.org/10.1007/s42161-020-00644-w) contains supplementary material, which is available to authorized users. * Awais Khan [email protected] 1
Plant Pathology and Plant-Microbe Biology Section, Cornell University, Geneva, NY 14456, USA
2
Eastern NY Commercial Horticulture Program, Cornell Cooperative Extension at Hudson Valley Research Lab, Highland, NY 12528, USA
(Stockwell et al. 2002). The bacteria enter the plants through natural openings or wounds in flowers, leaves, shoots, fruits, and rootstocks (Norelli et al. 2003; Vanneste 2000; Beer et al. 1996).
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