PCR amplification of the mitochondrial DNA minisatellite region to detect Schistosoma mansoni infection in Biomphalaria glabrata snails.
Schistosoma infection in Biomphalaria glabrata can be detected by either exposing snails to light to induce cercarial shedding or by squeezing them between glass slides to detect parasites in the digestive gland and other regions. The methods available are inefficient for identification of prepatent infections and do not allow the diagnosis of infection in snails that die before arriving in the laboratory. Furthermore, infection is undetectable after migration of sporocysts from the head-foot region of the snail. We examined the use of polymerase chain reaction (PCR) amplification of minisatellite repeats from Schistosoma mansoni mitochondrial DNA (mtDNA) to identify snail infection. We found that amplification of mtDNA under low stringency conditions (LS-PCR) allowed for the identification of specific S. mansoni infection in Biomphalaria snails. To confirm these results, specific amplification reactions were performed using 2 sets of primers that allowed for the diagnosis of infection and an internal control of the reaction (multiplex PCR). Results obtained using multiplex PCR demonstrated the ability of the assay to detect S. mansoni-specific infection. Thus, LS-PCR as well as specific multiplex PCR allow for the detection of prepatent infections and show high specificity for S. mansoni in comparison with other trematode infections in either living or dead snails.
Jannotti-Passos
L K
LK
Centro de Pesquisas René Rachou, Belo Horizonte, MG, Brazil.
Vidigal
T H
TH
Dias-Neto
E
E
Pena
S D
SD
Simpson
A J
AJ
Dutra
W O
WO
Souza
C P
CP
Carvalho-Parra
J F
JF
D000818:Animals; D001700:Biomphalaria / Q000469:parasitology*; D017931:DNA Primers / Q000235:genetics; D018509:DNA, Helminth / Q000032:analysis / Q000235:genetics; D004272:DNA, Mitochondrial / Q000032:analysis / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D018598:Minisatellite Repeats; D016133:Polymerase Chain Reaction; D012550:Schistosoma mansoni / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D016622:Silver Staining; D013045:Species Specificity
D017931:DNA Primers; D018509:DNA, Helminth; D004272:DNA, Mitochondrial