Molecular detection and identification of avian influenza viruses by cDNA microarray
| Author(s) | Maughan, Michele Nancy | |
| Date Accessioned | 2020-10-07T11:23:36Z | |
| Date Available | 2020-10-07T11:23:36Z | |
| Publication Date | 2006 | |
| Abstract | Avian influenza (AI) is a respiratory viral pathogen of major concern to poultry producers and public health officials across the world. Rapid detection and subtyping of influenza viruses is necessary in order to control outbreaks and maintain routine surveillance. Microarray technology is a relatively new means of detecting pathogens and characterizing their genomic content. An avian influenza virus-specific cDNA microarray has been created and shown to correctly detect and identify the H5, H7, and H9 hemagglutinin subtypes, the N1, N2, and N3 neuraminidase subtypes, and the matrix gene of AI. ☐ Our prototype AI cDNA microarray contains 16 elements representing the matrix, hemagglutinin, and neuraminidase genes of avian influenza isolates and a negative control from the F gene of Newcastle disease virus. These elements are spotted in duplicate in four subarrays yielding 8 spots per element and 128 spots total. To validate our microarray, an unknown panel comprised of 10 avian influenza isolates was tested. Of the ten isolates, 100% (10/10) were correctly identified as type A influenza viruses and 70% (7/10) were fully subtyped by their hemagglutinin and neuraminidase genes. Further characterization of 100% (4/4) of the H5 isolates was accomplished by quantifying hybridization signal strength between the extensive phylogenetic representation of the H5 hemagglutinin gene sequences on our microarray and the H5 isolates in the unknown panel. ☐ Our results demonstrate the ability of a cDNA microarray to detect, identify, subtype, and phylogenetically/geographically group various avian influenza isolates. Our method as validated here can identify type A influenza via the conserved matrix gene, differentiate between the H5, H7, and H9 hemagglutinin subtypes, and differentiate between the N1, N2, and N3 neuraminidase subtypes of avian influenza. Furthermore, our AI cDNA microarray demonstrates the ability to determine the phylogenetic/geographic group from which an H5 hemagglutinin subtype originates based on hybridization signal strength. ☐ This method can be applied to clinical situations pending further validation experiments to determine the sensitivity of the array and increasing the number of representative HA and NA subtypes on the array (i.e. H1-4, H6, H8, and H10-16 and N4-9). Also, other viral and/or bacterial pathogens could be added to the array to increase its diagnostic power and aid the medical community in differential diagnoses. | en_US |
| Advisor | Keeler, Calvin L., Jr. | |
| Degree | M.S. | |
| Department | University of Delaware, Department of Animal and Food Sciences | |
| Unique Identifier | 123440040 | |
| URL | https://udspace.udel.edu/handle/19716/27805 | |
| Publisher | University of Delaware | en_US |
| URI | https://login.udel.idm.oclc.org/login?url=https://www.proquest.com/docview/304859252?accountid=10457 | |
| dc.subject.lcsh | Avian influenza -- Diagnosis | |
| dc.subject.lcsh | DNA microarrays | |
| dc.subject.lcsh | Viruses -- Identification | |
| Title | Molecular detection and identification of avian influenza viruses by cDNA microarray | en_US |
| Type | Thesis | en_US |