Selection and versatile application of virus-specific aptamers

Zsófia Balogh, Gergely Lautner, Viola Bardóczy, Beata Komorowska, R. Gyurcsányi, T. Mészáros

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Although the significance of molecular diagnostics in routine plant virus detection is rapidly growing, the preferred methods are still antibody-based enzyme immunoassays. In the past decade, aptamers have been demonstrated to be viable alternatives of antibodies in many applications. We set out to select apple stem pitting virus (ASPV)-specific aptamers and to apply them as antibody substitutes in various immunoassay methods. The applied systematic evolution of ligands by exponential enrichment (SELEX) procedure resulted in highly discriminative aptamers selectively binding to the target virus coat protein even in complex protein matrixes. We developed protocols for exploitation of aptamers in diverse plant virus diagnosis methods, such as dot and Western blot analyses and enzyme-linked oligonucleotide assay (ELONA). Our selected aptamers proved to be superior to the available antibody in all aspects. In contrast to the antibody, the aptamers decorate both native and denaturated proteins, and ELONA produces higher signal intensity than traditional enzyme-linked immunosorbent assay (ELISA) with virus-infected plant extract. Summarily, our results present the selection and practical utilization of first plant virus-specific aptamers. Most important, the first application of ELONA for virus detection is demonstrated, which proposes a novel, more flexible, and cost-effective means of virus diagnostics.

Original languageEnglish
Pages (from-to)4187-4195
Number of pages9
JournalFASEB Journal
Volume24
Issue number11
DOIs
Publication statusPublished - Nov 2010

Fingerprint

Viruses
Plant Viruses
Antibodies
Oligonucleotides
Assays
Enzymes
SELEX Aptamer Technique
Molecular Pathology
Plant Extracts
Capsid Proteins
Malus
Immunoenzyme Techniques
Immunoassay
Proteins
Immunosorbents
Western Blotting
Enzyme-Linked Immunosorbent Assay
Costs and Cost Analysis
Pitting
Ligands

Keywords

  • DOS-ELONA
  • Immunoblot
  • SELEX
  • Virus coat protein

ASJC Scopus subject areas

  • Biochemistry
  • Biotechnology
  • Genetics
  • Molecular Biology

Cite this

Selection and versatile application of virus-specific aptamers. / Balogh, Zsófia; Lautner, Gergely; Bardóczy, Viola; Komorowska, Beata; Gyurcsányi, R.; Mészáros, T.

In: FASEB Journal, Vol. 24, No. 11, 11.2010, p. 4187-4195.

Research output: Contribution to journalArticle

Balogh, Zsófia ; Lautner, Gergely ; Bardóczy, Viola ; Komorowska, Beata ; Gyurcsányi, R. ; Mészáros, T. / Selection and versatile application of virus-specific aptamers. In: FASEB Journal. 2010 ; Vol. 24, No. 11. pp. 4187-4195.
@article{10ac332fabdc47af9f3d11c7fda39212,
title = "Selection and versatile application of virus-specific aptamers",
abstract = "Although the significance of molecular diagnostics in routine plant virus detection is rapidly growing, the preferred methods are still antibody-based enzyme immunoassays. In the past decade, aptamers have been demonstrated to be viable alternatives of antibodies in many applications. We set out to select apple stem pitting virus (ASPV)-specific aptamers and to apply them as antibody substitutes in various immunoassay methods. The applied systematic evolution of ligands by exponential enrichment (SELEX) procedure resulted in highly discriminative aptamers selectively binding to the target virus coat protein even in complex protein matrixes. We developed protocols for exploitation of aptamers in diverse plant virus diagnosis methods, such as dot and Western blot analyses and enzyme-linked oligonucleotide assay (ELONA). Our selected aptamers proved to be superior to the available antibody in all aspects. In contrast to the antibody, the aptamers decorate both native and denaturated proteins, and ELONA produces higher signal intensity than traditional enzyme-linked immunosorbent assay (ELISA) with virus-infected plant extract. Summarily, our results present the selection and practical utilization of first plant virus-specific aptamers. Most important, the first application of ELONA for virus detection is demonstrated, which proposes a novel, more flexible, and cost-effective means of virus diagnostics.",
keywords = "DOS-ELONA, Immunoblot, SELEX, Virus coat protein",
author = "Zs{\'o}fia Balogh and Gergely Lautner and Viola Bard{\'o}czy and Beata Komorowska and R. Gyurcs{\'a}nyi and T. M{\'e}sz{\'a}ros",
year = "2010",
month = "11",
doi = "10.1096/fj.09-144246",
language = "English",
volume = "24",
pages = "4187--4195",
journal = "FASEB Journal",
issn = "0892-6638",
publisher = "FASEB",
number = "11",

}

TY - JOUR

T1 - Selection and versatile application of virus-specific aptamers

AU - Balogh, Zsófia

AU - Lautner, Gergely

AU - Bardóczy, Viola

AU - Komorowska, Beata

AU - Gyurcsányi, R.

AU - Mészáros, T.

PY - 2010/11

Y1 - 2010/11

N2 - Although the significance of molecular diagnostics in routine plant virus detection is rapidly growing, the preferred methods are still antibody-based enzyme immunoassays. In the past decade, aptamers have been demonstrated to be viable alternatives of antibodies in many applications. We set out to select apple stem pitting virus (ASPV)-specific aptamers and to apply them as antibody substitutes in various immunoassay methods. The applied systematic evolution of ligands by exponential enrichment (SELEX) procedure resulted in highly discriminative aptamers selectively binding to the target virus coat protein even in complex protein matrixes. We developed protocols for exploitation of aptamers in diverse plant virus diagnosis methods, such as dot and Western blot analyses and enzyme-linked oligonucleotide assay (ELONA). Our selected aptamers proved to be superior to the available antibody in all aspects. In contrast to the antibody, the aptamers decorate both native and denaturated proteins, and ELONA produces higher signal intensity than traditional enzyme-linked immunosorbent assay (ELISA) with virus-infected plant extract. Summarily, our results present the selection and practical utilization of first plant virus-specific aptamers. Most important, the first application of ELONA for virus detection is demonstrated, which proposes a novel, more flexible, and cost-effective means of virus diagnostics.

AB - Although the significance of molecular diagnostics in routine plant virus detection is rapidly growing, the preferred methods are still antibody-based enzyme immunoassays. In the past decade, aptamers have been demonstrated to be viable alternatives of antibodies in many applications. We set out to select apple stem pitting virus (ASPV)-specific aptamers and to apply them as antibody substitutes in various immunoassay methods. The applied systematic evolution of ligands by exponential enrichment (SELEX) procedure resulted in highly discriminative aptamers selectively binding to the target virus coat protein even in complex protein matrixes. We developed protocols for exploitation of aptamers in diverse plant virus diagnosis methods, such as dot and Western blot analyses and enzyme-linked oligonucleotide assay (ELONA). Our selected aptamers proved to be superior to the available antibody in all aspects. In contrast to the antibody, the aptamers decorate both native and denaturated proteins, and ELONA produces higher signal intensity than traditional enzyme-linked immunosorbent assay (ELISA) with virus-infected plant extract. Summarily, our results present the selection and practical utilization of first plant virus-specific aptamers. Most important, the first application of ELONA for virus detection is demonstrated, which proposes a novel, more flexible, and cost-effective means of virus diagnostics.

KW - DOS-ELONA

KW - Immunoblot

KW - SELEX

KW - Virus coat protein

UR - http://www.scopus.com/inward/record.url?scp=78649852048&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=78649852048&partnerID=8YFLogxK

U2 - 10.1096/fj.09-144246

DO - 10.1096/fj.09-144246

M3 - Article

C2 - 20624933

AN - SCOPUS:78649852048

VL - 24

SP - 4187

EP - 4195

JO - FASEB Journal

JF - FASEB Journal

SN - 0892-6638

IS - 11

ER -