title: Molecular differentiation of Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis by pyrosequencing.
abstract: Nematodes of the genus Trichinella which infect wildlife and domestic animals show a cosmopolitan distribution. These zoonotic parasites are the aetiological agents of a severe human disease, trichinellosis. Twelve taxa are recognized in the Trichinella genus, but they cannot be identified by morphology since they are sibling species/genotypes. For epidemiological studies, it is extremely important to identify each taxon since they have different distribution areas and host ranges. In the present study, polymerase chain reaction (PCR) amplification of the mitochondrial large subunit ribosomal RNA (lsu-RNA) gene coupled with a pyrosequencing technique was developed to distinguish among four Trichinella species: Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis. A PCR method was used to amplify the lsu-RNA of Trichinella sp. larvae in mouse muscles and single larvae collected from infected muscles by digestion. The results show that the four Trichinella species can be distinguished by using 26 nucleotides in the target region and the method is sensitive enough to identify individual larvae. The pyrosequencing provides a simple, rapid and high-throughput tool for the differentiation of Trichinella species.
authors.affiliation: Department of Parasitology,Faculty of Medicine, Khon Kaen University,Khon Kaen40002,Thailand.
authors.forename: L
authors.initials: L
authors.lastname: Sadaow
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University,Khon Kaen40002,Thailand.
authors.forename: C
authors.initials: C
authors.lastname: Tantrawatpan
authors.affiliation: Department of Parasitology,Faculty of Medicine, Khon Kaen University,Khon Kaen40002,Thailand.
authors.forename: P M
authors.initials: PM
authors.lastname: Intapan
authors.affiliation: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University,Khon Kaen40002,Thailand.
authors.forename: V
authors.initials: V
authors.lastname: Lulitanond
authors.affiliation: Department of Parasitology,Faculty of Medicine, Khon Kaen University,Khon Kaen40002,Thailand.
authors.forename: T
authors.initials: T
authors.lastname: Boonmars
authors.affiliation: Department of Parasitology,Faculty of Medicine, Chiang Mai University,Chiang Mai50200,Thailand.
authors.forename: N
authors.initials: N
authors.lastname: Morakote
authors.affiliation: Department of Infectious, Parasitic and Immunomediated Diseases,Istituto Superiore di Sanità,viale Regina Elena 299,00161Rome,Italy.
authors.forename: E
authors.initials: E
authors.lastname: Pozio
authors.affiliation: Department of Parasitology,Faculty of Medicine, Khon Kaen University,Khon Kaen40002,Thailand.
authors.forename: W
authors.initials: W
authors.lastname: Maleewong
mesh_terms: D000818:Animals; D001483:Base Sequence; D018509:DNA, Helminth / Q000235:genetics; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D058889:Molecular Typing / Q000379:methods*; D013552:Swine; D013553:Swine Diseases / Q000469:parasitology*; D014234:Trichinella / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D017160:Trichinella spiralis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D014235:Trichinellosis / Q000469:parasitology* / Q000662:veterinary*
chemical_list: D018509:DNA, Helminth