Characterization of rDNA sequences from Syphacia obvelata, Syphacia muris, and Aspiculuris tetraptera and development of a PCR-based method for identification

• Published in: Veterinary parasitology 2008-05, Vol.153 (3), p.379-383
• Main Authors: Parel, Joan Dee C., Galula, Jedhan U., Ooi, Hong-Kean
• Format: Article
• Language: English
• Subjects: animal morphology; Animals; Animals, Wild - parasitology; Aspiculuris tetraptera; Base Sequence; DNA, Helminth - chemistry; DNA, Helminth - genetics; DNA, Ribosomal Spacer - chemistry; DNA, Ribosomal Spacer - genetics; Enterobiasis - diagnosis; Enterobiasis - parasitology; Enterobiasis - veterinary; Enterobius - genetics; Enterobius - isolation & purification; Gene Amplification; ITS-2; Mice; Molecular Sequence Data; nematode infections; nucleotide sequences; Pinworms; polymerase chain reaction; Polymorphism, Restriction Fragment Length; Rats; rDNA; ribosomal DNA; Rodent Diseases - diagnosis; Rodent Diseases - parasitology; Sequence Alignment - veterinary; Species Specificity; Syphacia muris; Syphacia obvelata
• ISSN: 0304-4017; 1873-2550
• DOI: 10.1016/j.vetpar.2008.02.001

To differentiate the morphologically similar pinworms of the common laboratory rodents, such as Syphacia obvelata and Syphacia muris, we amplified and sequenced the region spanning the internal transcribed spacer 1 (ITS-1), 5.8S gene, and ITS-2 of the ribosomal DNA followed by designing of species-specific primers for future use in the identification of the worms. It was observed that S. obvelata, S. muris and Aspiculuris tetraptera can be differentiated from each other based on their rDNA sequences. This is the first report of the ITS-1, 5.8S, and ITS-2 of the rDNA of the three aforementioned rodent pinworm species. The use of restriction endonucleases, AluI or RsaI, further allowed the delineation of the three species. Moreover, we also constructed species-specific primers that were designed for unique regions of the ITS-2 of the three species. This approach allowed their specific identification with no amplicons being amplified from heterogenous DNA samples, and sequencing confirmed the identity of the sequences amplified. Thus, the use of these specific primers along with PCR-RFLP can serve as useful tools for the identification of pinworms in rats, mice, and wild rodents.