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

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.

Авторы
Razo S.C. 1, 2 , Panferov V.G.1 , Safenkova I.V.1 , Varitsev Y.A.3 , Zherdev A.V. 1 , Pakina E.N. 2 , Dzantiev B.B. 1
Журнал
Издательство
MDPI AG
Номер выпуска
11
Язык
Английский
Статус
Опубликовано
Том
18
Год
2018
Организации
  • 1 A.N. Bach Institute of Biochemistry, Research Centre of Biotechnology of the Russian Academy of Sciences, Leninsky Prospect 33, Moscow, 119071, Russian Federation
  • 2 RUDN University, Miklukho-Maklaya Street 8/2, Moscow, 117198, Russian Federation
  • 3 A.G. Lorch All-Russian Potato Research Institute, Kraskovo, Lorch Street 23, 140051 Moscow, Russia. varyuriy@yandex.ru
Ключевые слова
gold nanoparticles; increase of sensitivity; lateral flow test strips; potato virus Y; pre-incubation; sandwich immunoassay
Дата создания
04.02.2019
Дата изменения
04.02.2019
Постоянная ссылка
https://repository.rudn.ru/ru/records/article/record/36238/
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