Lateral flow immunoassay for rapid detection of potato ring rot caused by Clavibacter michiganensis subsp. sepedonicus

Safenkova, Irina V.; Zaitsev, I. A.; Pankratova, Galina; Varitsev, Yuri A.; Zherdev, Anatoly V.; Dzantiev, Boris B.

doi:10.1134/s0003683814120011

Cited by 11 publications

(5 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Their use in phytopathology was focused mostly on fungi [ 18 , 19 ] and viruses [ 20 , 21 ], although it has been extended to the specific detection of bacteria such as Erwinia amylovora [ 22 ], Clavibacter michiganensis subsp. sepedonicus [ 23 ], and Xanthomonas campestris pv. musacearum [ 24 ].…”

Section: Introductionmentioning

confidence: 99%

Lateral flow immunoassay for on-site detection of Xanthomonas arboricola pv. pruni in symptomatic field samples

et al. 2017

View full text Add to dashboard Cite

Xanthomonas arboricola pv. pruni is a quarantine pathogen and the causal agent of the bacterial spot disease of stone fruits and almond, a major threat to Prunus species. Rapid and specific detection methods are essential to improve disease management, and therefore a prototype of a lateral flow immunoassay (LFIA) was designed for the detection of X. arboricola pv. pruni in symptomatic field samples. It was developed by producing polyclonal antibodies which were then combined with carbon nanoparticles and assembled on nitrocellulose strips. The specificity of the LFIA was tested against 87 X. arboricola pv. pruni strains from different countries worldwide, 47 strains of other Xanthomonas species and 14 strains representing other bacterial genera. All X. arboricola pv. pruni strains were detected and cross-reactions were observed only with four strains of X. arboricola pv. corylina, a hazelnut pathogen that does not share habitat with X. arboricola pv. pruni. The sensitivity of the LFIA was assessed with suspensions from pure cultures of three X. arboricola pv. pruni strains and with spiked leaf extracts prepared from four hosts inoculated with this pathogen (almond, apricot, Japanese plum and peach). The limit of detection observed with both pure cultures and spiked samples was 104 CFU ml-1. To demonstrate the accuracy of the test, 205 samples naturally infected with X. arboricola pv. pruni and 113 samples collected from healthy plants of several different Prunus species were analyzed with the LFIA. Results were compared with those obtained by plate isolation and real time PCR and a high correlation was found among techniques. Therefore, we propose this LFIA as a screening tool that allows a rapid and reliable diagnosis of X. arboricola pv. pruni in symptomatic plants.

show abstract

Section: Introductionmentioning

confidence: 99%

Lateral flow immunoassay for on-site detection of Xanthomonas arboricola pv. pruni in symptomatic field samples

et al. 2017

View full text Add to dashboard Cite

show abstract

“…The conjugates were synthesized as described in Safenkova et al, 15 . The antibodies were dialyzed against a 1,000-fold volume of 10-mM Tris-HCl, pH 9.0, at +4 °C for 12 h. The pH of the GNP solution was adjusted to 9.0 with 0.2 M of K 2 CO 3 .…”

Section: Synthesis Of Gnp-antibody Conjugatesmentioning

confidence: 99%

“…Membrane compounds and multimembrane composite were prepared using approaches described in Safenkova et al 15 . The GNP-antibody conjugate was deposited onto CNPC-12µ membranes from a solution that had optical density equal to 10 at λ= 520 nm.…”

Section: Preparation Of Test Stripsmentioning

confidence: 99%

“…Currently, the list of agricultural bacteria detected by commercially available LFIA systems is small. Similarly, developments described in the literature are limited 6,10,[14][15][16] . Therefore, considering the number of serious bacterial infections known to affect crops and the associated losses 17,18 , the development of LFIA tests for bacterial phytopathogens is a high priority.…”

mentioning

confidence: 99%

“…All reactants are applied to the test strip before the assay, and contact between the strip and the test sample initiates specific interactions that yield a visible result: the presence or absence of coloration on certain areas of the strip. Sample preparation consists only of simple, rapid homogenization of the tissue in the extracting solution [11][12][13][14][15] . Currently, the list of agricultural bacteria detected by commercially available LFIA systems is small.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Lateral Flow Immunoassay for Rapid Diagnosis of Potato Blackleg Caused by Pectobacterium atrosepticum

Safenkova¹

2015

Biosci., Biotech. Res. Asia

View full text Add to dashboard Cite

Potato blackleg and soft rot caused by Pectobacterium atrosepticum lead to significant yield losses. The early detection of P. atrosepticum is essential for healthy potato seed tubers. The aim of this study was to develop a method to rapidly detect P. atrosepticum based on the lateral flow immunoassay (LFIA) technique. Rabbit polyclonal antibodies specific to various strains of P. atrosepticum were obtained. Conjugates of these antibodies with gold nanoparticles averaging 20 nm in diameter were synthesized. Optimal concentrations of antibodies and conjugates deposited on membranes of the test strips were determined. The developed LFIA is suitable for analyzing potato tubers and leaves and has a visual detection limit of 2 × 10 5 cells/mL and a duration time of 10 min. Simple, rapid preparation of samples consists of homogenization in extracting buffer. No cross-reactivities with other potato pathogens, such as Pectobacterium carotovorum subsp. carotovorum and Dickeya dianthicola, or saprophytes of healthy potato plants were detected. The assay was tested on 30 lots of potato tubers. The LFIA results were confirmed by ELISA (100% concurrence) and PCR (87.5% for positive samples and 95.5% for negative samples). Diagnosis of potato blackleg and soft rot by LFIA requires no equipment or training to perform, is cost effective and can be used in the field to monitor infection-causing P. atrosepticum.

show abstract

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

Safenkova

Zaitsev²,

Varitsev³

et al. 2016

Anal Bioanal Chem

View full text Add to dashboard Cite

Early detection of potato infections is essential for effective disease management. The aim of this study was to develop a lateral flow immunoassay (LFIA) for rapid detection of a serious potato disease, potato blackleg, caused by Dickeya dianthicola and Dickeya solani. Polyclonal antibodies specific to different strains of Dickeya were obtained from rabbits after immunization with bacterial cells of D. dianthicola and D. solani. Enzyme-linked immunosorbent assay testing with use of a wide range of bacterial species showed that the polyclonal antibodies detect closely related strains of D. dianthicola and D. solani. Cross-reactivity with widespread pathogenic bacteria (nine species) and saprophytes of healthy potato plants was not detected. The LFIA based on the obtained antibodies and gold nanoparticles with average diameter of 20 nm was developed. Under optimized conditions, the LFIA method enabled the analysis of potato extracts within 10 min, with a visual limit of detection of 1 × 10 CFU/ml for leaves and 4 × 10 CFU/ml for tubers. The assay was tested on potato stem and tuber extracts, and the results of the LFIA were confirmed in 92.1% of samples using the real-time polymerase chain reaction. The findings confirmed that the developed LFIA could be used for monitoring blackleg infection without the need for special equipment or skills. Graphical Abstract The developed lateral flow immunoassay is an efficient tool for rapid detection of a serious potato disease, potato blackleg, caused by Dickeya dianthicola and Dickeya solani.

show abstract

Lateral flow immunoassay for rapid detection of potato ring rot caused by Clavibacter michiganensis subsp. sepedonicus

Cited by 11 publications

References 22 publications

Lateral flow immunoassay for on-site detection of Xanthomonas arboricola pv. pruni in symptomatic field samples

Lateral flow immunoassay for on-site detection of Xanthomonas arboricola pv. pruni in symptomatic field samples

Lateral Flow Immunoassay for Rapid Diagnosis of Potato Blackleg Caused by Pectobacterium atrosepticum

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

Contact Info

Product

Resources

About