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Molecular differentiation of Trichinella spiralis, T. pseudospiralis, T. papuae and T. zimbabwensis by pyrosequencing

Published online by Cambridge University Press:  13 May 2013

L. Sadaow
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand
C. Tantrawatpan
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand Division of Cell Biology, Department of Preclinical Sciences, Faculty of Medicine, Thammasat University, Rangsit Campus, Pathum Thani12121, Thailand
P.M. Intapan
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand
V. Lulitanond
Affiliation:
Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand
T. Boonmars
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand
N. Morakote
Affiliation:
Department of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai50200, Thailand
E. Pozio
Affiliation:
Department of Infectious, Parasitic and Immunomediated Diseases, Istituto Superiore di Sanità, viale Regina Elena 299, 00161Rome, Italy
W. Maleewong*
Affiliation:
Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen40002, Thailand
*

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.

Type
Short Communications
Copyright
Copyright © Cambridge University Press 2013 

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