Characterization of Oesophagostomum spp. from pigs in China by PCR-based approaches using genetic markers in the internal transcribed spacers of ribosomal DNA

Abstract

In the present study, samples of Oesophagostomum spp. collected from pigs from different geographical localities in mainland China were characterized genetically by polymerase chain reaction-linked single-strand conformation polymorphism (PCR-SSCP) and restriction fragment length polymorphism (PCR-RFLP) techniques using genetic markers in the internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The second internal transcribed spacer (ITS-2) was amplified from 51 individual nodule worms by PCR, and the amplicons were analyzed by SSCP. With the exception of slight microheterogeneity, SSCP analyses displayed two distinct banding profiles that allowed the identification of all Oesophagostomum spp. samples examined into two groups, the first one represented O. dentatum, and the second one may represent O. quadrispinulatum. Then, the entire ITS was amplified from individual samples, and the amplicons were digested with restriction endonuclease Pst I. The results of RFLP analyses were consistent with that of SSCP. Sequence analysis of ITS rDNA supported the identification and differentiation of Chinese Oesophagostomum spp. samples into two species, namely, O. dentatum and O. quadrispinulatum. These PCR-based approaches provide useful complementary tools to traditional methods for the accurate identification of Oesophagostomum spp. (irrespective of developmental stage) and have implications for studying the ecology and population genetic structures of these parasites and for the prevention and control of the diseases they cause.