Protocol for DNA extraction of Cryptosporidium spp. oocysts in fecal samples.
Molecular characterization of Cryptosporidium spp.oocysts in clinical samples is useful for public health since it allows the study of sources of contamination as well as the transmission in different geographical regions. Although widely used in developed countries, in Brazil it is restricted to academic studies, mostly using commercial kits for the extraction of genomic DNA, or in collaboration with external reference centers, rendering the method expensive and limited. The study proposes the application of the modifications recently introduced in the method improving feasibility with lower cost. This method was efficient for clinical samples preserved at -20 degrees C for up to six years and the low number of oocysts may be overcome by repetitions of extraction.
Serviço de Parasitologia da Divisão de Laboratório Central, Hospital das Clínicas, Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil.
Elenice M N
EM
Gonçalves
Ronalda S
RS
Araújo
Magali
M
Orban
Glavur R
GR
Matté
Maria Helena
MH
Matté
Carlos E P
CE
Corbett
D000818:Animals; D002985:Clinical Protocols; D003457:Cryptosporidiosis / Q000175:diagnosis*; D003458:Cryptosporidium / Q000235:genetics* / Q000302:isolation & purification; D016054:DNA, Protozoan / Q000302:isolation & purification*; D005243:Feces / Q000469:parasitology*; D006801:Humans; D033661:Oocysts; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D016054:DNA, Protozoan