title: Differentiation of Cucumber mosaic virus isolates by hybridization to oligonucleotides in a microarray format.
abstract: A system for microarrays was developed to detect and differentiate Cucumber mosaic virus (CMV) serogroups and subgroups. The coat protein genes of 14 different isolates were amplified using cy3-labelled generic but species-specific primers. These amplicons were hybridized against a set of five different serotype and subgroup specific 24-mer oligonucleotides bound to an aldehyde-coated glass slide via an aminolinker. The results of the hybridization revealed that the method allowed a clear differentiation of the 14 different CMV isolates into the serogroupes 1 and 2, and in addition was able to assign 9 out of 10 different serogroup 1 isolates correctly into subgroups 1a and 1b. This differentiation was not possible by RFLP analysis with the restriction enzyme MspI. The use of amplicons larger than 700 base pairs and their successful differentiation by hybridization to specific oligonucleotides opens avenues to highly parallel, yet sensitive assays for plant viruses.
authors.affiliation: Biocentre Klein Flottbek, Department of Phytomedicine, University of Hamburg, Ohnhorststr. 18, D22609 Hamburg, Germany.
authors.forename: Zhang
authors.initials: Z
authors.lastname: Deyong
authors.forename: Peter
authors.initials: P
authors.lastname: Willingmann
authors.forename: Cornelia
authors.initials: C
authors.lastname: Heinze
authors.forename: Günter
authors.initials: G
authors.lastname: Adam
authors.forename: Monica
authors.initials: M
authors.lastname: Pfunder
authors.forename: Beatrice
authors.initials: B
authors.lastname: Frey
authors.forename: Jürg E
authors.initials: JE
authors.lastname: Frey
mesh_terms: D036022:Capsid Proteins / Q000235:genetics; D017799:Cucumovirus / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D012367:RNA, Viral / Q000235:genetics
chemical_list: D036022:Capsid Proteins; D015345:Oligonucleotide Probes; D009841:Oligonucleotides; D012367:RNA, Viral