Journal article
Rapid detection of avian influenza virus in chicken fecal samples by immunomagnetic capture reverse transcriptase–polymerase chain reaction assay
Section of Poultry Diseases, Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark1
Division of Poultry, Fish and Fur Animals, National Veterinary Institute, Technical University of Denmark2
National Veterinary Institute, Technical University of Denmark3
Section for Veterinary Diagnostics, Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark4
Division of Veterinary Diagnostics and Research, National Veterinary Institute, Technical University of Denmark5
BioLabChip Group, LabChip Section, Department of Micro- and Nanotechnology, Technical University of Denmark6
LabChip Section, Department of Micro- and Nanotechnology, Technical University of Denmark7
Department of Micro- and Nanotechnology, Technical University of Denmark8
Avian influenza virus (AIV) causes great economic losses for the poultry industry worldwide and threatens the human population with a pandemic. The conventional detection method for AIV involves sample preparation of viral RNA extraction and purification from raw sample such as bird droppings. In this study, magnetic beads were applied for immunoseparation and purification of AIV from spiked chicken fecal sample.
The beads were conjugated with monoclonal antibodies against the AIV nucleoprotein, which is conserved in all the AIV. The bead-captured virus was detected by reverse transcriptase–polymerase chain reaction (RT-PCR) without RNA extraction because of effective removal of RT-PCR inhibitors. The developed bead-based assay showed a similar detection limit comparable to the RNA extraction and the classic virus isolation method.
Using ready-to-use antibody-conjugated bead, the method requires less than 5 h. Furthermore, the method has potential to integrate into a Lab-on-a-chip system for rapid detection and identification of AIV.
| Language: | English |
|---|---|
| Year: | 2011 |
| Pages: | 258-265 |
| ISSN: | 18790070 and 07328893 |
| Types: | Journal article |
| DOI: | 10.1016/j.diagmicrobio.2010.09.022 |
| ORCIDs: | Yi, Sun , Wolff, Anders and Bang, Dang Duong |
Agglutination Tests Animals Antibodies, Monoclonal Antibodies, Viral Chickens Feces Immunomagnetic Separation Influenza A virus Influenza in Birds NP protein, Influenza A virus Nucleocapsid Proteins Poultry Diseases RNA, Viral RNA-Binding Proteins Reverse Transcriptase Polymerase Chain Reaction Sensitivity and Specificity Viral Core Proteins
