Evaluation of the specificity of five oligoprobes for identification of cyathostomin species from horses.
Here, we report evaluation of five oligoprobes designed from intergenic spacer (IGS) region sequences for identification of cyathostomin species. Oligoprobes were designed for identification of Cylicocyclus ashworthi, Cylicocyclus nassatus, Cylicostephanus longibursatus, Cylicostephanus goldi and a fifth probe designed to identify all members of this tribe. PCR amplification of IGS DNA from 16 cyathostomin species allowed sequence comparison and identification of four putative species-specific probes. Southern blotting of amplified products from 16 species showed that all probes were species-specific. The fifth probe recognised all 16 cyathostomin species but did not bind to members of the genus Strongylus. Furthermore, these probes were used to identify individual infective L3, eggs and L4 indicating that they will be invaluable to furthering the study of the epidemiology and pathogenesis of these important equine nematodes.
Division of Equine Studies, Department of Veterinary Clinical Sciences and Animal Husbandry, University of Liverpool, Leahurst, South, CH64 7TE, Wirral, UK. jhodgkin@liverpool.ac.uk
J E
JE
Hodgkinson
S
S
Love
J R
JR
Lichtenfels
S
S
Palfreman
Y H
YH
Ramsey
J B
JB
Matthews
D000818:Animals; D001483:Base Sequence; D018509:DNA, Helminth / Q000032:analysis / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000032:analysis / Q000235:genetics*; D006734:Horse Diseases / Q000469:parasitology*; D006736:Horses; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction / Q000379:methods; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013045:Species Specificity; D017206:Strongylida Infections / Q000469:parasitology / Q000662:veterinary*; D013320:Strongyloidea / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification
D018509:DNA, Helminth; D021903:DNA, Ribosomal Spacer; D015345:Oligonucleotide Probes