Polymerase chain reaction and restriction fragment length polymorphism analysis of the ITS2 region for differentiation of Brazilian Biomphalaria intermediate hosts of the Schistosoma mansoni.
We sequenced the internal transcribed spacer 2 of the ribosomal DNA (ITS2-DNAr) from the three Schistosoma mansoni intermediate hosts in Brazil: Biomphalaria glabrata, Biomphalaria tenagophila and Biomphalaria straminea. Analysis of a restriction map from those sequences allowed us to select putative restriction enzymes able to identify the snail species under study. Four restriction enzymes were used and HpaII provided simple species-specific profiles easily visualized in polyacrylamide gels. The use of ITS2 is advantageous as it provides a small fragment of 460 bp which may be easily amplified by PCR. In the current work, we showed that the amplification of ITS2-DNAr together with HpaII enzyme restriction is an auxiliary molecular tool for the morphological identification of such snails as well as for taxonomic and phylogenetic studies of neotropical planorbids.
Vidigal
Teofânia Heloísa Dutra Amorim
TH
Centro de Pesquisas René Rachou, Fundação Oswaldo Cruz, Belo Horizonte, MG, Brasil.
Magalhães
Kelly Grace
KG
Carvalho Od
Omar dos Santos
Odos S
D000818:Animals; D001700:Biomphalaria / Q000145:classification / Q000201:enzymology / Q000235:genetics*; D004262:DNA Restriction Enzymes / Q000032:analysis; D004275:DNA, Ribosomal / Q000032:analysis; D021903:DNA, Ribosomal Spacer / Q000032:analysis; D004199:Disease Vectors; D004591:Electrophoresis, Polyacrylamide Gel; D005819:Genetic Markers; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012550:Schistosoma mansoni
D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer; D005819:Genetic Markers; D004262:DNA Restriction Enzymes