Development of a DNA microarray for simultaneous detection and genotyping of lyssaviruses.
The lyssavirus genus of the Rhabdoviridae family of viruses includes 7 genotypes and several non-assigned isolates. The source of lyssavirus infections is diverse with numerous reservoirs in a wide geographical area. In many parts of the world reservoir hosts can potentially be carrying one of several lyssavirus strains and possibly new divergent isolates await discovery. Accordingly, generic detection methods are required to be able to detect and discriminate all lyssaviruses and identify new divergent isolates. Here we have allied a sequence-independent amplification method to microarray to enable simultaneous detection and identification of all lyssavirus genotypes. To do so, lyssavirus RNA was converted to cDNA and amplified in a random PCR, labelled and hybridized to probes on the microarray chip before being statistically analysed. The probes were to a 405 bp region of the relatively conserved N gene. Here we demonstrate a microarray capable of detecting each of the seven lyssavirus genotypes. The random amplification of lyssavirus RNA and the numerous oligonucleotide probes on the microarray chip also offer the potential to detect novel lyssaviruses.
Division of Microbial Sciences, Faculty of Health and Medical Sciences, University of Surrey, Guildford, Surrey, GU2 7XH, United Kingdom.
R
R
Gurrala
A
A
Dastjerdi
N
N
Johnson
J
J
Nunez-Garcia
S
S
Grierson
F
F
Steinbach
M
M
Banks
D000818:Animals; D000835:Animals, Wild / Q000821:virology; D018076:DNA, Complementary / Q000235:genetics; D018114:Lyssavirus / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018353:Rhabdoviridae Infections / Q000662:veterinary; D014764:Viral Proteins / Q000235:genetics
D018076:DNA, Complementary; D012367:RNA, Viral; D014764:Viral Proteins