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DNA microarray technique for detection and identification of seven Flaviviruses pathogenic for man

Nordstrom, H. (author)
Swedish Institute for Infectious Disease Control, Solna, Sweden
Falk, K. I. (author)
Karolinska Institutet
Lindegren, G. (author)
Swedish Institute for Infectious Disease Control, Solna, Sweden
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Mouzavi-Jazi, M. (author)
Swedish Institute for Infectious Disease Control, Solna, Sweden
Walden, A. (author)
Royal Institute of Technology, Department of Biotechnology, Stockholm, Sweden
Elgh, Fredrik (author)
Umeå universitet,Patologi,Virologi
Nilsson, Peter (author)
KTH,Proteomik
Lundkvist, A. (author)
Karolinska Institutet
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 (creator_code:org_t)
2005
2005
English.
In: Journal of Medical Virology. - New York : Wiley. - 0146-6615 .- 1096-9071. ; 77:4, s. 528-540
  • Journal article (peer-reviewed)
Abstract Subject headings
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  • A flavivirus microarray was developed for detection and identification of yellow fever (YF), West Nile, Japanese encephalitis (JE), and the dengue 1-4 viruses, which are causing severe human disease all over the world. The microarray was based on 500-nucleotide probe fragments from five different parts of the seven viral genomes. A low-stringent amplification method targeting the corresponding regions of the viral genomic RNA was developed and combined with hybridization to the microarray for detection and identification. For distinction of the generated virus-specific fluorescence-patterns a fitting analysis procedure was adapted. The method was verified as functional for all seven flaviviruses and the strategy for the amplification, combined with the long probes, provided a high tolerance for smaller genetic variability, most suitable for these rapidly changing RNA viruses. A potentially high detection and identification capacity was proven on diverged strains of West Nile and dengue viruses. The lower limit for detection was equivalent, or better, when compared to routinely used RT-PCR methods. The performance of the method was verified on human patient samples containing dengue viruses, or normal human serum spiked with YF or JE viruses. The results demonstrated the ability of the flavivirus microarray to screen simultaneously a sample for several viruses in parallel, in combination with a good lower limit of detection.

Keyword

emerging viruses
new strains
simultaneous screening
mismatch tolerance
west-nile-virus
reverse-transcriptase pcr
polymerase chain-reaction
dengue virus
phylogenetic-relationships
rapid identification
influenza-viruses
yellow-fever
population
hybridization

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ref (subject category)
art (subject category)

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