DETECTION OF SCHISTOSOMA-MANSONI IN BIOMPHALARIA USING NESTED PCR

A nested polymerase chain reaction (PCR) protocol was developed for de tecting the presence of Schistosoma mansoni sporocysts in intermediate host snails of the genus Biomphalaria. To accomplish this, rDNA genes encoding the 18S rRNA of S. mansoni and Biomphalaria alexandrina from Egypt were sequenced, as were 18S-encoding genes of the 13-16-R1 and Salvador strains of Biomphalaria glabrata. Based on a comparison of ho st and parasite sequences, a nested set of PCR primers was designed to allow specific amplification of portions of S. mansoni 18S rDNA. Thes e primers allowed detection of as little as 10 fg of S. mansoni DNA di luted in 100 ng of snail DNA and did not allow amplification of snail 18S sequences. Using nested PCR, the presence of a single S. mansoni s porocyst within an adult snail could be detected at 1 day postexposure . In DNA samples extracted from each of 74 snails of the M-line strain of B. glabrata exposed to from 1 to 10 S. mansoni miracidia for inter vals ranging from 1 to 44 days, use of the outside primer pair alone d etected the parasite's presence in 51% of the snails, whereas the sequ ential use of outside and nested primer pairs detected parasites in 92 % of the snails. This approach has utility in determining if snails in endemic areas bear prepatent or inactive infections and in assessing the degree of compatibility between local snail and schistosome popula tions. It will also facilitate studies of resistance of snails to infe ction.