Analysis of Schistosoma mansoni population structure using total fecal egg sampling.
Many parasite populations are difficult to sample because they are not uniformly distributed between several host species and are often not easily collected from the living host, thereby limiting sample size and possibly distorting the representation of the population. For the parasite Schistosoma mansoni, we investigated the use of eggs, in aggregate, from the stools of infected individuals as a simple and representative sample. Previously, we demonstrated that microsatellite allele frequencies can be accurately estimated from pooled DNA of cloned S. mansoni adults. Here, we show that genotyping of parasite populations from reproductively isolated laboratory strains can be used to identify these specific populations based on characteristic patterns of allele frequencies, as observed by polyacrylamide gel electrophoresis and automated sequencer analysis of fluorescently labeled PCR products. Microsatellites used to genotype aggregates of eggs collected from stools of infected individuals produced results consistent with the geographic distribution of the samples. Preferential amplification of smaller alleles, and stutter PCR products, had negligible effect on measurement of genetic differentiation. Direct analysis of total stool eggs can be an important approach to questions of population genetics for this parasite by increasing the sample size to thousands per infected individual and by reducing bias.
Blank
Walter A
WA
Center for Global Health and Diseases, Case Western Reserve University, Cleveland, Ohio 44106, USA.
Reis
Eliana A G
EA
Thiong'o
Fred W
FW
Braghiroli
Joao F
JF
Santos
Jarbas M
JM
Melo
Paulo R S
PR
Guimarães
Isabel C S
IC
Silva
Luciano K
LK
Carmo
Theomira M A
TM
Reis
Mitermeyer G
MG
Blanton
Ronald E
RE
D000818:Animals; D001938:Brazil; D018509:DNA, Helminth / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D005243:Feces / Q000469:parasitology*; D005260:Female; D005787:Gene Frequency; D005838:Genotype; D006801:Humans; D007630:Kenya; D008297:Male; D018895:Microsatellite Repeats / Q000235:genetics*; D010063:Ovum; D016133:Polymerase Chain Reaction; D012550:Schistosoma mansoni / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012555:Schistosomiasis mansoni / Q000469:parasitology*; D017421:Sequence Analysis
D018509:DNA, Helminth