title: A Multiplex-PCR approach to identification of the Brazilian intermediate hosts of Schistosoma mansoni.
abstract: Due to difficulties concerning morphological identification of planorbid snails of the genus Biomphalaria, and given a high variation of characters and in the organs with muscular tissue, we designed specific polymerase chain reaction (PCR) primers for Brazilian snail hosts of Schistosoma mansoni from available sequences of internal transcribed spacer 2 (ITS2) of the ribosomal RNA gene. From the previous sequencing of the ITS2 region, one primer was designed to anchor in the 5.8S conserved region and three other species-specific primers in the 28S region, flanking the ITS2 region. These four primers were simultaneously used in the same reaction (Multiplex-PCR), under high stringency conditions. Amplification of the ITS2 region of Biomphalaria snails produced distinct profiles (between 280 and 350 bp) for B. glabrata, B. tenagophila and B. straminea. The present study demonstrates that Multiplex-PCR of ITS2-DNAr showed to be a promising auxiliary tool for the morphological identification of Biomphalaria snails, the intermediate hosts of S. mansoni.
authors.affiliation: Departamento de Zoologia, Instituto de Ciências Biológicas, UFMG, Belo Horizonte, MG, Brasil.
authors.forename: T H D A
authors.initials: TH
authors.lastname: Vidigal
authors.forename: K G
authors.initials: KG
authors.lastname: Magalhães
authors.forename: J C
authors.initials: JC
authors.lastname: Kissinger
authors.forename: R L
authors.initials: RL
authors.lastname: Caldeira
authors.forename: A J G
authors.initials: AJ
authors.lastname: Simpson
authors.forename: O S
authors.initials: OS
authors.lastname: Carvalho
mesh_terms: D000818:Animals; D001700:Biomphalaria / Q000145:classification / Q000235:genetics*; D001938:Brazil; D017931:DNA Primers; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D004199:Disease Vectors; D016133:Polymerase Chain Reaction / Q000379:methods*; D012550:Schistosoma mansoni / Q000302:isolation & purification*; D012552:Schistosomiasis / Q000517:prevention & control; D016622:Silver Staining; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D021903:DNA, Ribosomal Spacer