Rapid PCR-based delineation of the porcine nodular worms, Oesophagostomum dentatum and O. quadrispinulatum.
At some stages of development, it is impossible to identify the porcine nodular worms Oesophagostomum dentatum and O. quadrispinulatum to the species level using morphological parameters. A molecular approach utilizing genetic markers in the second internal transcribed spacer (ITS-2) of ribosomal (r) DNA was developed to overcome this limitation. The ITS-2 sequence of each species was determined, and specific oligonucleotide primers were designed to regions of greatest sequence difference between the species. Utilizing these primers, rapid PCR procedures were developed for the specific amplification of DNA of O. dentatum or O. quadrispinulatum, which are now used routinely to monitor the purity of larval cultures and to confirm the identity of larvae derived from the intestine or faeces. The application of specific PCR has major implications for studying the population biology of nodular worms in the pig model.
Newton
L A
LA
Department of Veterinary Science, University of Melbourne, Werribee, Victoria, Australia.
Chilton
N B
NB
Monti
J R
JR
Bjørn
H
H
Várady
M
M
Christensen
C M
CM
Gasser
R B
RB
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D018509:DNA, Helminth / Q000032:analysis*; D004275:DNA, Ribosomal; D005819:Genetic Markers; D007814:Larva; D008969:Molecular Sequence Data; D009814:Oesophagostomiasis / Q000469:parasitology; D009815:Oesophagostomum / Q000145:classification / Q000235:genetics*; D016133:Polymerase Chain Reaction; D013045:Species Specificity; D013552:Swine
D017931:DNA Primers; D018509:DNA, Helminth; D004275:DNA, Ribosomal; D005819:Genetic Markers