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 porcin e nodular worms Oesophagostomum dentatum and 0. quadrispinulatum to th e species level using morphological parameters. A molecular approach u tilizing genetic markers in the second internal transcribed spacer (IT S-2) of ribosomal (r) DNA was developed to overcome this limitation. T he ITS-2 sequence of each species was determined, and specific oligonu cleotide primers were designed to regions of greatest sequence differe nce between the species. Utilizing these primers, rapid PCR procedures were developed for the specific amplification of DNA of 0. dentatum o r 0. quadrispinulatum, which are now used routinely to monitor the pur ity of larval cultures and to confirm the identity of larvae derived f rom the intestine or faeces. The application of specific PCR has major implications for studying the population biology of nodular worms in the pig model. (C) 1997 Academic Press Limited.