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High throughput sequencing of the Angiostrongylus cantonensis genome: a parasite spreading worldwide

Published online by Cambridge University Press:  17 July 2013

ALESSANDRA L. MORASSUTTI*
Affiliation:
Laboratório de Biologia Parasitária da Faculdade de Biociências e Laboratório de Parasitologia Molecular do Instituto de Pesquisas Biomédicas da Pontifícia Universidade do Rio Grande do Sul (PUCRS), Av Ipiranga 6690, 90690-900 Porto Alegre RS, Brasil
ANDREY PERELYGIN
Affiliation:
Division of Parasitic Diseases & Malaria, Center for Global Health, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Atlanta, GA, 30329, USA
MARCOS O. DE CARVALHO
Affiliation:
Programa de Pós-Graduação em Genética e Biologia Molecular, Universidade Federal do Rio Grande do Sul, Porto Alegre RS, Brasil
LEANDRO NASCIMENTO LEMOS
Affiliation:
Universidade Federal do Pampa – Campus São Gabriel, Av. Antônio Trilha, 1847 – São Gabriel – RS – CEP: 97300-000
PAULO MARCOS PINTO
Affiliation:
Universidade Federal do Pampa – Campus São Gabriel, Av. Antônio Trilha, 1847 – São Gabriel – RS – CEP: 97300-000
MIKE FRACE
Affiliation:
Division of Parasitic Diseases & Malaria, Center for Global Health, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Atlanta, GA, 30329, USA
PATRICIA P. WILKINS
Affiliation:
Division of Parasitic Diseases & Malaria, Center for Global Health, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Atlanta, GA, 30329, USA
CARLOS GRAEFF-TEIXEIRA
Affiliation:
Laboratório de Biologia Parasitária da Faculdade de Biociências e Laboratório de Parasitologia Molecular do Instituto de Pesquisas Biomédicas da Pontifícia Universidade do Rio Grande do Sul (PUCRS), Av Ipiranga 6690, 90690-900 Porto Alegre RS, Brasil
ALEXANDRE J. DA SILVA
Affiliation:
Division of Parasitic Diseases & Malaria, Center for Global Health, Centers for Disease Control and Prevention, 1600 Clifton Road NE, Atlanta, GA, 30329, USA
*
*Corresponding author. Instituto de Pesquisas Biomédicas da PUCRS, Avenida Ipiranga 6690, 2 andar, Sala 20, CEP: 90690-900 Porto Alegre RS, Brazil. E-mail: [email protected]

Summary

Angiostrongylus cantonensis is a parasitic nematode of rodents and a leading aetiological agent of eosinophilic meningitis in humans. Definitive diagnosis is difficult, often relying on immunodiagnostic methods which utilize crude antigens. New immunodiagnostic methods based on recombinant proteins are being developed, and ideally these methods would be made available worldwide. Identification of diagnostic targets, as well as studies on the biology of the parasite, are limited by a lack of molecular information on Angiostrongylus spp. available in databases. In this study we present data collected from DNA random high-throughput sequencing together with proteomic analyses and a cDNA walking methodology to identify and obtain the nucleotide or amino acid sequences of unknown immunoreactive proteins. 28 080 putative ORFs were obtained, of which 3371 had homology to other deposited protein sequences. Using the A. cantonensis genomic sequences, 156 putative ORFs, matching peptide sequences obtained from previous proteomic studies, were considered novel, with no homology to existing sequences. Full-length coding sequences of eight antigenic target proteins were obtained. In this study we generated not only the complete nucleotide sequences of the antigenic protein targets but also a large amount of genomic data which may help facilitate future genomic, proteomic, transcriptomic or metabolomic studies on Angiostrongylus.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2013 

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