Development of a Nano-ELISA system for the rapid and sensitive detection of H9N2 avian influenza

Document Type: Original Articles

Authors

1 Department of Biology, Faculty of Basic science, Payame Noor University, Tehran, Iran

2 Proteomics and Biochemistry Department, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.

3 Department of Microbiology, School of Specialized Veterinary Science, Science and Research Branch, Islamic Azad University, Tehran, Iran.

4 Department of Proteomics and Biochemistry, Razi vaccine and Serum Research Institute, Karaj, Iran

5 Department of medical Nanotechnology, school of advanced Medical Technology, Tehran University of Medical Sciences, Tehran, Iran

6 Department of cellular and molecular Biology, Faculty of biological sciences, Kharazmi University, Tehran, Iran

Abstract

Influenza is one of the most important viral diseases that is common among the birds and the mammals
and is caused by specific viruses that belong to the Orthomyxoviridae family. Migratory aquatic birds are
the reservoir of the disease and there is a likelihood of the disease in any region. There are different
methods for detecting the avian influenza, but by the point of detection rates, the ELISA may be one of
the most important current methods. In this work we synthesized Gold nanoparticles and conjugated it
with rabbit-anti-chicken IgG-HRP. An ELISA test was done to compare the bioactivity of Au-antichicken
HRP with anti-chicken HRP in order to detect the antibody against the H9N2 subtype of avian
influenza virus. Using 133 field chicken sera, the sensitivity of nano-ELISA as compared to traditional
ELISA was calculated to be 100%, whereas the specificity was 92%. This method was significantly more
sensitive than the traditional ELISA and didn’t require extra costs. It can therefore be concluded that the
AuNP-HRP conjugate can be applicable in immune analysis procedure where a more confident result is
required.

Keywords

Main Subjects


Article Title [French]

Développement d’un système Nano-ELISA pour la détection rapide et sensible du virus d’influenza aviaire H9N2

Abstract [French]

L’influenza est l’une des maladies virales les plus importantes et les plus répandues chez les oiseaux et les mammifères. Un virus appartenant à la famille des Orthomyxoviridae est à l’origine de cette pathologie. Les oiseaux migratoires aquatiques représentent le réservoir naturel de ce virus et cette maladie peut donc affecter toutes les régions du monde. Il existe différentes méthodes de détection de l’influenza aviaire, mais l’ELISA semble représenter l’une des méthodes les plus pertinentes avec un fort taux de détection. Dans cette étude, nous avons synthétisé des nanoparticules d’or associées à des anticorps de lapin anti-IgG de poulet marqués à l’HRP. Un test ELISA a été effectué afin de comparer la bio-activité des anticorps associés et non-associés aux particules d’or dans la détection du sous-type H9N2 du virus de l’influenza aviaire. La sensibilité de notre nano-ELISA a été comparée à celle de l’ELISA traditionnel sur 133 sérums de poulets fermiers. Les résultats obtenus montrent que la sensibilité du nano-ELISA est supérieure à l’ELISA traditionnel (6 échantillons positifs supplémentaires détectés) avec une spécificité de 92%. Etant donnée la sensibilité plus importante des anticorps de lapin associés aux nanoparticules d’or, l’utilisation de ces réactifs peut augmenter la fiabilité des essais immunologiques diagnostiques.

Keywords [French]

  • Influenza aviaire
  • ELISA
  • nanoparticule d’or

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