Fecal specimens preparation methods for PCR diagnosis of human taeniosis.
Sample preparation and DNA extraction protocols for DNA amplification by PCR, which can be applied in human fecal samples for taeniasis diagnosis, are described. DNA extracted from fecal specimens with phenol/chloroform/isoamilic alcohol and DNAzol reagent had to be first purified to generate fragments of 170 pb and 600 pb by HDP2-PCR. This purification step was not necessary with the use of QIAmp DNA stool mini kit. Best DNA extraction results were achieved after eggs disruption with glass beads, either with phenol/chloroform/isoamilic alcohol, DNAzol reagent or QIAmp DNA stool mini kit.
Departamento de Apoio, Produção e Saúde Animal, FOA, UNESP, Araçatuba, SP, Brazil. caris@fmva.unesp.br
Cáris Maroni
CM
Nunes
Luiz Gustavo Ferraz
LG
Lima
Camila Santos
CS
Manoel
Rodrigo Norberto
RN
Pereira
Mauro Massaharu
MM
Nakano
José Fernando
JF
Garcia
D000818:Animals; D018509:DNA, Helminth / Q000737:chemistry* / Q000302:isolation & purification; D004587:Electrophoresis, Agar Gel; D005243:Feces / Q000469:parasitology*; D006801:Humans; D016133:Polymerase Chain Reaction; D013045:Species Specificity; D013048:Specimen Handling / Q000379:methods*; D041202:Taenia saginata / Q000145:classification / Q000235:genetics*; D041201:Taenia solium / Q000145:classification / Q000235:genetics*; D013622:Taeniasis / Q000175:diagnosis* / Q000469:parasitology
D018509:DNA, Helminth