Multiarray on a test strip (MATS): rapid multiplex immunodetection of priority potato pathogens

Abstract

Multiarray on a test strip (MATS) was developed for the detection of eight important potato pathogens. The proposed assay combines the rapidity of immunochromatography with the high throughput of array techniques. The test zone of the immunochromatographic strip comprises ordered rows of spots containing antibodies specific for different potato pathogens. The assay benefits from the simplicity of immunochromatography; colored immune complexes form at the corresponding spots within the test zone. The presence and intensity of the coloration are used for identification of the target pathogens. The MATS was applied to the simultaneous detection of eight priority potato pathogens, characterized by the following limits of detection: 1 ng/mL for potato virus X and the ordinary type of potato virus Y, 10 ng/mL for potato virus M, 20 ng/mL for potato leaf roll virus, 40 ng/mL for necrotic-type potato virus Y, 100 ng/mL for potato virus S, 300 ng/mL for potato virus A, and 10(4) cells/mL for Clavibacter michiganensis subsp. sepedonicus. Analysis time was 15 min. The observed sensitivity of the MATS was comparable to the traditional enzyme-linked immunosorbent assay. The developed technique was tested on potato leaf extracts, and its efficiency for on-site control of the pathogens was confirmed in 100 % by commercial LFIA test strips. Graphical abstract Location of binding zones in the developed multiarray on a test strip (MATS) for simultaneous detection of eight pathogens.

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

“…The ELISA test of the antibody and antibody–GNP conjugates was carried out according to Safenkova et al [ 34 ] with the following modifications: PVY N at a concentration of 0.5 μg∙mL −1 was immobilized in microplate wells at 37 °C for 2 h; after the washing steps with PBST, anti-PVY antibodies at concentrations ranging from 0.45 ng∙mL −1 to 1000 ng∙mL −1 (or serial dilutions of the GNP–antibody conjugates from 1:10 to 2:10 4 ) were used. After the completion of all of the stages, including the addition of the anti-rabbit antibody–peroxidase conjugate, the TMB substrate, and the stop solution (1 M H 2 SO 4 ), the optical density was measured at 450 nm (OD 450 ).…”

Ways to Reach Lower Detection Limits of Lateral Flow Immunoassays

“…Examples of test systems based on the principle of "two-dimensional immunochromatography" are presented in the works of Taranova et al [104] on the detection of drugs and Safenkova et al [148] on the detection of phytopathogens. General approaches to multizonal LFIA were discussed in Hu et al [149], and the current state of the development of multiplex immunoassays was discussed in Li et al [150].…”

Silver-enhanced lateral flow immunoassay for highly-sensitive detection of potato leafroll virus

“…PLRV was propagated in Datura stramonium and purified from infected leaves (Rowhani & Stacesmith, 1979). Polyclonal antibodies (fraction of IgG) specific to PLRV described by Safenkova et al (2016) were used in the experiments. Tris(hydroxymethyl)aminomethane (Tris), Triton X-100, 3,3 ′ ,5,5 ′ -tetramethylbenzidine dihydrochloride (TMB), sodium azide, biotinamidohexanoic acid N-hydroxysuccinimide ester, dimethyl sulfoxide (DMSO), and chloroauric acid were purchased from Fluka (Germany); staphylococcal protein A was obtained from Imtek (Russia); streptavidin-polyperoxidase conjugate, bovine serum albumin (BSA), sodium citrate, silver lactate, hydroquinone, and sucrose were obtained from Sigma-Aldrich (USA); glycerol, NaCl, and K 2 CO 3 were obtained from DiaM (Russia); and Na 2 CO 3 , NaHCO 3 , KH 2 PO 4 , and KOH were obtained from Khimmed (Russia).…”