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MOLECULAR DIFFERENTIATION OF ANGIOSTRONGYLUS COSTARICENSIS, A. CANTONENSIS, AND A. VASORUM BY POLYMERASE CHAIN REACTION- RESTRICTION FRAGMENT LENGTH POLYMORPHISM
Caldeira, Roberta Lima et al. | Date Issued: 2003
Author
Caldeira, Roberta Lima
Carvalho, Omar dos Santos
Mendonça, Cristiane Lafetá Furtado de
Teixeira, Carlos Graeff
Silva, Márcia C. F.
Ben, Renata
Maurer, Rafael
Lima, Walter S.
Lenzi, Henrique Leonel
Affilliation
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintoses Intestinais. Belo Horizonte, MG, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintoses Intestinais. Belo Horizonte, MG, Brasil
Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Helmintoses Intestinais. Belo Horizonte, MG, Brasil
Pontifícia Universidade Católica do Rio Grande do Sul. Laboratório de Parasitologia Molecular e Biologia de Parasitas. Porto Alegre, RS, Brasil
Pontifícia Universidade Católica do Rio Grande do Sul. Laboratório de Parasitologia Molecular e Biologia de Parasitas. Porto Alegre, RS, Brasil
Pontifícia Universidade Católica do Rio Grande do Sul. Laboratório de Parasitologia Molecular e Biologia de Parasitas. Porto Alegre, RS, Brasil
Pontifícia Universidade Católica do Rio Grande do Sul. Laboratório de Parasitologia Molecular e Biologia de Parasitas. Porto Alegre, RS, Brasil
Universidade Federal de Minas Gerais.Instituto de Ciências Biológicas. Departamento de Parasitologia. Belo Horizonte, MG, Brasil
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.
Abstract
Angiostrongylus cantonensis, A. costaricensis, and A. vasorum are etiologic agents of human parasitic diseases. Their identification, at present, is only possible by examining the adult worm after a 40-day period following infection of vertebrate hosts with the third-stage larvae. In order to obtain a diagnostic tool to differentiate larvae and adult worm from the three referred species, polymerase chain reaction-restriction fragment length polymorphism was carried out. The rDNA second internal transcribed spacer (ITS2) and mtDNA cytochrome oxidase I regions were amplified, followed by digestion of fragments with the restriction enzymes RsaI, HapII, AluI, HaeIII, DdeI and ClaI. The enzymes RsaI and ClaI exhibited the most discriminating profiles for the differentiation of the regions COI of mtDNA and ITS2 of rDNA respectively. The methodology using such regions proved to be efficient for the specific differentiation of the three species of Angiostrongylus under study.
Keywords
Angiostrongylus
Polymerase chain reaction
Restriction fragment length polymorphism
Second internal transcribed spacer
Cytochrome oxidase I
 
DeCS
Angiostrongylus
Reação em Cadeia da Polimerase
Polimorfismo de Fragmento de Restrição
Citocromos
 
Publisher
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz
Citation
CALDEIRA, Roberta L. et al. Molecular differentiation of Angiostrongylus costaricensis, A. cantonensis, and A. vasorum by polymerase chain reaction-restriction fragment length polymorphism. Memórias do Instituto Oswaldo Cruz, Rio de Janeiro, v. 98, n. 8, p. 1039-1043, 2003.
DOI
10.1590/S0074-02762003000800011
ISSN
0074-0276
Notes
Acknowledgements to Dr Kentaro Yoshimura, Department of Parasitology, Medical School of University of Akita, Japan, for providing us with Angiostrongylus cantonensis; Joziana Barçante, Department of Parasitology of Federal University of Minas Gerais, Brazil, for maintaining the life cycle of A. vasorum; Ester Maria Mota, Department of Pathology, Oswaldo Cruz Institute-Fiocruz, Rio de Janeiro, Brazil, for being in charge of A. costaricensis life cycle.