Multiplex PCR for both identification of Brazilian Biomphalaria species (Gastropoda: Planorbidae) and diagnosis of infection by Schistosoma mansoni (Trematoda: Schistosomatidae).
A simple and single-step technique based on multiplex PCR (multiplex polymerase chain reaction) has been developed for simultaneous identification of Brazilian Biomphalaria species, the intermediate hosts of Schistosoma mansoni, and their diagnosis of infection by the trematode. We used species-specific primers directed both to the internal transcribed spacer 2 (ITS2) of ribosomal DNA from 3 of the S. mansoni host species and to the mitochondrial DNA (mtDNA) from the trematode. Those primers were used simultaneously in a single multiplex-PCR reaction, and template DNA was obtained from S. mansoni-infected and noninfected snails. The results were visualized in silver stained polyacrylamide gels, revealing the presence of specific bands. The methodology has shown to be efficient, fast, and reproducible for Biomphalaria species identification and diagnosis of snails infected by S. mansoni during prepatent periods.
Jannotti-Passos
Liana K
LK
Centro de Pesquisas René Rachou-Fiocruz, Av. Augusto de Lima 1715, 30190-002 Belo Horizonte, MG, Brazil. passoslk@cpqrr.fiocruz.br
Magalhães
Kelly Grace
KG
Carvalho
Omar S
OS
Vidigal
Teofânia H D A
TH
D000818:Animals; D001700:Biomphalaria / Q000145:classification* / Q000235:genetics / Q000469:parasitology*; D001938:Brazil; D017931:DNA Primers / Q000737:chemistry; D004272:DNA, Mitochondrial / Q000032:analysis; D021903:DNA, Ribosomal Spacer / Q000032:analysis; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D015203:Reproducibility of Results; D012550:Schistosoma mansoni / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D004272:DNA, Mitochondrial; D021903:DNA, Ribosomal Spacer; D012337:RNA, Ribosomal, 18S