Molecular differentiation of Angiostrongylus costaricensis, A. cantonensis, and A. vasorum by polymerase chain reaction-restriction fragment length polymorphism.
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.
Caldeira
Roberta L
RL
Laboratório de Helmintoses Intestinais, Centro de Pesquisas René Rachou-Fiocruz, Av. Augusto de Lima 1715, 30190-002 Belo Horizonte, MG, Brasil.
Carvalho
Omar S
OS
Mendonça
Cristiane L
CL
Graeff-Teixeira
Carlos
C
Silva
Márcia C
MC
Ben
Renata
R
Maurer
Rafael
R
Lima
Walter S
WS
Lenzi
Henrique L
HL
D000801:Angiostrongylus / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D000818:Animals; D004262:DNA Restriction Enzymes / Q000032:analysis; D004272:DNA, Mitochondrial / Q000032:analysis; D005819:Genetic Markers; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012335:RNA, Ribosomal / Q000032:analysis; D013045:Species Specificity
D004272:DNA, Mitochondrial; D005819:Genetic Markers; D012335:RNA, Ribosomal; D004262:DNA Restriction Enzymes