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Detection and subtyping of influenza A virus based on a short oligonucleotide microarray
- Source :
-
Diagnostic Microbiology & Infectious Disease . Nov2009, Vol. 65 Issue 3, p261-270. 10p. - Publication Year :
- 2009
-
Abstract
- Abstract: We report the design and characterization of a microarray with 46 short virus-specific oligonucleotides for detecting influenza A virus of 5 subtypes: H1N1, H1N2, H3N2, H5N1, and H9N2. A unique combination of 3 specific modifications was introduced into the microarray assay: (1) short probes of 19 to 27 nucleotides, (2) simple amplification of full-length hemagglutinin and neuraminidase cDNAs with universal primers, and (3) Klenow-mediated labeling and further amplification of the samples before hybridization. The assay correctly and specifically detected and subtyped 11 different influenza A isolates from human, avian, and swine species representing the 5 subtypes. When tested with 225 clinical samples, 20 were detected to be positive using our microarray-based assay, whereas only 10 were positive by the conventional culture method. The entire analysis was completed within 7 h. Thus, these modifications result in a specific, sensitive, and rapid microarray assay and may be used for constructing microarrays for the detection of all influenza subtypes and strains. [Copyright &y& Elsevier]
Details
- Language :
- English
- ISSN :
- 07328893
- Volume :
- 65
- Issue :
- 3
- Database :
- Academic Search Index
- Journal :
- Diagnostic Microbiology & Infectious Disease
- Publication Type :
- Academic Journal
- Accession number :
- 44582522
- Full Text :
- https://doi.org/10.1016/j.diagmicrobio.2009.07.016