Development of a lateral flow immunoassay for rapid diagnosis of potato blackleg caused by Dickeya species

Safenkova, Irina V.; Zaitsev, I. A.; Varitsev, Yuri A.; Byzova, Nadezhda A.; Drenova, Natalia V.; Zherdev, Anatoly V.; Dzantiev, Boris B.

doi:10.1007/s00216-016-0140-6

Cited by 18 publications

(5 citation statements)

References 39 publications

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“…The homogeneity of the GNPs in the solution and the absence of aggregates was shown by dynamic light scattering (DLS) (data shown in ESM, Figure 5 Sc), which also confirmed the dimensions of the nanoparticles (the average hydrodynamic diameter was 19.5 nm). Based on our previous experiences in the synthesis of GNP conjugates, we confirmed that spherical GNPs with diameters of 20–30 nm provided high stability to the conjugate [ 36 , 37 ]. Based on the characterized GNPs, we synthesized a GNP conjugate with the anti-PVY antibodies.…”

Section: Resultssupporting

confidence: 64%

How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

Razo

Panferov

Safenkova

et al. 2018

Sensors

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A simple approach was proposed to decrease the detection limit of sandwich lateral flow immunoassay (LFIA) by changing the conditions for binding between a polyvalent antigen and a conjugate of gold nanoparticles (GNPs) with antibodies. In this study, the potato virus Y (PVY) was used as the polyvalent antigen, which affects economically important plants in the Solanaceae family. The obtained polyclonal antibodies that are specific to PVY were characterized using a sandwich enzyme-linked immunosorbent assay (ELISA) and surface plasmon resonance (SPR). For LFIA, the antibodies were conjugated with GNPs with a diameter of 17.4 ± 1.0 nm. We conducted LFIAs using GNP conjugates in a dried state on the test strip and after pre-incubation with a sample. Pre-incubating the GNP conjugates and sample for 30 s was found to decrease the detection limit by 60-fold from 330 ng∙mL−1 to 5.4 ng∙mL−1 in comparison with conventional LFIA. The developed method was successfully tested for its ability to detect PVY in infected and uninfected potato leaves. The quantitative results of the proposed LFIA with pre-incubation were confirmed by ELISA, and resulted in a correlation coefficient of 0.891. The proposed approach is rapid, simple, and preserves the main advantages of LFIA as a non-laboratory diagnostic method.

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Section: Resultssupporting

confidence: 64%

How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

Razo

Panferov

Safenkova

et al. 2018

Sensors

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“…Dickeya species can be identified using a series of biochemical tests paired with semi-selective media as well as lateral flow immunoassay [17]; neither method is time efficient or accurate. In contrast, molecular techniques are specific and sensitive for detection and differentiation of Dickeya to species [18–21].…”

Section: Introductionmentioning

confidence: 99%

Development of a robust, field-deployable loop-mediated isothermal amplification (LAMP) assay for specific detection of potato pathogen Dickeya dianthicola targeting a unique genomic region

et al. 2019

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Destructive maceration, a wide host range, and longevity in non-plant substrates has established Dickeya dianthicola (blackleg of potato) as a significant threat to potato industries worldwide. To protect these businesses, a specific and sensitive point-of-care D . dianthicola detection tool is necessary. We have developed a loop-mediated isothermal amplification (LAMP) assay for specific, sensitive, and rapid detection of D . dianthicola , which can be streamlined for point-of-care use. The developed LAMP assay targets a unique gene, alcohol dehydrogenase , of D . dianthicola . Assay specificity was assessed using strains present in inclusivity (16 D . dianthicola strains) and exclusivity panels (56 closely related, potato pathogenic, and other bacterial strains). Amplification with strains of inclusivity panel occurred, and cross-reactivity with non-target DNA was not observed. The limit of detection (LOD) was 10 CFU/ml when dilutions were made before isolating the genomic DNA; however, LOD was determined as 1 pg using 10-fold serially diluted D . dianthicola genomic DNA. Similar LOD of 1 pg was observed when serially diluted target genomic DNA was mixed with host genomic DNA. LOD (1 pg) was also calculated with 10-fold serially diluted synthetic DNA fragments containing primer target sites. Naturally and artificially inoculated plant samples were used for field adaptability tests with the field-deployable Optigene Plant Material Lysis Kit and a heat block (65°C); the results were obtained within 20 minutes. Despite the lack of method precision, no false positives or false negatives were observed. Therefore, with prepared reactions and a steady heat source, this assay can be used for rapid point-of-care detection, which is imperative for quarantine, eradication, disease management, and border protection.

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“…As shown in Table 3 , the performance of our sensing platform is competitive with respect to other recent technologies in terms of sensitivity and the limit of detection. The two techniques based on lateral flow immune assays offer the great advantage to be very cheap and simple, two key aspects for on field analysis, but on the other hand they are not quantitative [ 24 , 25 ]. Papadakis and coworkers proposed an approach based on quartz crystal microbalance to detect DNA amplificons and they showed a LOD of 10 3 CFU/mL that is lower than our devices but their approach requires additional steps for DNA extraction from plant tissues and DNA amplification [ 26 ], which results in an increase in the cost and time necessary for the analysis.…”

Section: Discussionmentioning

confidence: 99%

Pectobacterium atrosepticum Biosensor for Monitoring Blackleg and Soft Rot Disease of Potato

et al. 2020

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Pectobacterium atrosepticum (Pba) is a quarantine and threatening phytopathogen known as the causal agent of blackleg and soft rot disease of potatoes in many areas. Its early detection is then important to have healthy potato tubers and reduce economic losses. Today, conventional methods such as enzyme-linked immunosorbent-assay (ELISA) and polymerase chain reaction (PCR) are typically used for Pba detection, but they are expensive and time-consuming. Here we report on the optimization of an alternative approach based on an electrochemical impedance immunosensor combining a microfluidic module and a microelectrodes array, and having advantages in terms of low cost, ease of use and portability. For validation and for assessing its performance, the lab-on-chip platform has been compared with two standard methods (ELISA and PCR).

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Development of a lateral flow immunoassay for rapid diagnosis of potato blackleg caused by Dickeya species

Cited by 18 publications

References 39 publications

How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

How to Improve Sensitivity of Sandwich Lateral Flow Immunoassay for Corpuscular Antigens on the Example of Potato Virus Y?

Development of a robust, field-deployable loop-mediated isothermal amplification (LAMP) assay for specific detection of potato pathogen Dickeya dianthicola targeting a unique genomic region

Pectobacterium atrosepticum Biosensor for Monitoring Blackleg and Soft Rot Disease of Potato

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