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- Title
Development of species-specific primers for the ectoparasitic nematode speciesXiphinema brevicolle, X. diffusum, X. elongatum, X. ifacolumandX. longicaudatum(Nematoda: Longidoridae) based on ribosomal DNA sequences.
- Authors
Oliveira, Claudio M.; Fenton, Brian; Malloch, Gaynor; Brown, Derek J. F.; Neilson, Roy
- Abstract
The objective of this study was to develop single-step PCR species-specific primers that reliably discriminate four economically importantXiphinemaspecies (X. brevicolle, X. elongatum, X. ifacolumandX. longicaudatum) andX. diffusumthat is taxonomically very similar toX. brevicolle.Each species-specific reverse primer was located in the ITS-1 rDNA region and was used in combination with a universal forward primer located in the 18S rDNA gene. Primer reliability was confirmed by screening seven and 11 populations, respectively of X.diffusumandX. elongatum.Potential species-specific primers were also identified forX. brevicolle, X. longicaudatumandX. ifacolum, however too few populations of these species were available to thoroughly assess their reliability. For all species-specific primers, specificity was demonstrated by the absence of cross-reactions with 14 non-targetXiphinemaspecies. Multiplex PCR was effective and reproducible for two (X. longicaudatumandX. ifacolum) or three (X. brevicolle, X. diffusumandX. elongatum) of the target nematode species, thus improving the applicability of the diagnostic primers.
- Subjects
POLYMERASE chain reaction; DAGGER nematodes; RECOMBINANT DNA; NUCLEOTIDE sequence; GENES
- Publication
Annals of Applied Biology, 2005, Vol 146, Issue 3, p281
- ISSN
0003-4746
- Publication type
Article
- DOI
10.1111/j.1744-7348.2005.040031.x