Development and laboratory evaluation of a polymerase chain reaction for monitoring Schistosoma mansoni infestation of water.
A sensitive and specific detection of cercariae of human schistosomes is required for better definition of risk of infection. In the present study, we have developed a polymerase chain reaction (PCR) assay for the detection of cercariae of Schistosoma mansoni in water. A simple DNA preparation was adapted for this purpose, and PCR primers were designed based on the 121-basepair highly repeated sequence we previously identified in the genome of S. mansoni. The PCR assay detected as little as 10(-6) ng of S. mansoni DNA, and the high sensitivity enabled the detection of a single cercaria. For trapping of cercariae we adapted a filtration apparatus previously used for separating schistosome eggs from turbid enzymatic digests of tissues. A single cercaria could be detected in repeated tests of water filtrates. Since the target DNA is tandemly arranged, a ladder pattern of the PCR products was demonstrated. A direct relationship was demonstrated between the number of ladder bands of the amplification products, and DNA concentration or number of cercariae. The feasibility of semiquantitation of schistosome larvae in natural water was thus suggested. The potential of the procedures described here for epidemiologic studies is discussed.
Hamburger
J
J
Hebrew University, Hadassah Medical School, Jerusalem Israel.
Xu
Y X
YX
Ramzy
R M
RM
Jourdane
J
J
Ruppel
A
A
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers / Q000737:chemistry; D018509:DNA, Helminth / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D005618:Fresh Water / Q000469:parasitology*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012550:Schistosoma mansoni / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013045:Species Specificity
D017931:DNA Primers; D018509:DNA, Helminth