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Reliable determination of hookworm nematode parasite species from dogs can be carried out post-mortem by microscopical examination of the buccal cavity of adult worms isolated from the host small intestine. In order to allow a proper diagnosis of the hookworm infection in a living host, we developed a polymerase chain reaction (PCR). Using DNA extracted from hookworm eggs and an oligonucleotide set designed on the basis of the DNA sequence of a cysteine proteinase AcCP1 gene, Ancylostoma caninum and Uncinaria stenocephala DNA can be discriminated from each other.