Development of molecular approaches for the identification of transmission sites of schistosomiasis.
Primers targeting the gene encoding the small subunit rRNA were designed to amplify DNA from Schistosoma mansoni with high specificity. Three PCR systems were developed: conventional PCR, two-step nested PCR (NPCR) and single-tube nested PCR (STNPCR). The limits of detection of parasite DNA for the conventional PCR, NPCR and STNPCR were 10 pg, 0.1 fg and 1 fg, respectively. The assays were highly specific for S. mansoni and did not recognise DNA from closely related non-schistosome trematodes. Using pools of Biomphalaria molluscs, PCR, NPCR and STNPCR were positive in 6/16 (37.5%), 15/16 (93.8%) and 13/16 (81.3%) of the tested samples, respectively, whereas the observation of cercariae shedding after exposure to light was able to detect S. mansoni infection in 6/16 (37.5%) of the pools. Thus, the molecular detection systems had a higher level of sensitivity than standard screening of intermediate hosts by cercarial shedding when DNA was purified from pools of snails collected from endemic areas. These PCR protocols have potential to be used as tools for monitoring of schistosome transmission.
Melo
Fábio L
FL
Departamento de Imunologia, Centro de Pesquisas Aggeu Magalhães-FIOCRUZ, Av. Prof. Moraes Rego s/n, Cidade Universitária, 50670-420, Recife, Brazil.
Gomes
Ana Lisa do Vale
AL
Barbosa
Constança S
CS
Werkhauser
Roberto P
RP
Abath
Frederico G C
FG
D000818:Animals; D001700:Biomphalaria / Q000469:parasitology*; D018509:DNA, Helminth / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards; D012550:Schistosoma mansoni / Q000469:parasitology*; D012552:Schistosomiasis / Q000469:parasitology* / Q000635:transmission; D012680:Sensitivity and Specificity
D018509:DNA, Helminth