[Purification and breaking techniques for cysts of Giardia spp].
The purpose of this study was to optimize and evaluate the purification techniques, isolation and breaking of cysts of Giardia spp from fecal samples to isolate DNA. Filtrated fecal samples were tested in 3 purification techniques: Telleman solution, sucrose and Telleman plus sucrose. The sucrose solution let us to isolate the cysts with less detritus. The cleaned cysts were splited in 3 techniques to test the breaking: osmotic shock and heat, chemistry degradation and thermic shock, enzymatic action and mechanic effect. Only the last method was successful and showed bands in agarose gel. The result of this study shows a routine and common method which could be used in the previous steps to the PCR technique for the genotypification of these parasites.
Polverino
D
D
Fac. de Cs. Médicas, Universidad Nacional de La Plata, Av 60 y 120 s/n, 1900 La Plata, Buenos Aires.
Molina
N B
NB
Minvielle
M C
MC
Lozano
M E
ME
Basualdo
J A
JA
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002469:Cell Separation / Q000379:methods*; D016054:DNA, Protozoan / Q000302:isolation & purification; D004285:Dogs; D004587:Electrophoresis, Agar Gel; D019286:Endopeptidase K / Q000494:pharmacology; D005243:Feces / Q000469:parasitology*; D005872:Giardia / Q000166:cytology / Q000235:genetics / Q000302:isolation & purification*; D006358:Hot Temperature; D006801:Humans; D033661:Oocysts / Q000737:chemistry / Q000187:drug effects / Q000302:isolation & purification*; D009997:Osmotic Pressure; D012965:Sodium Chloride / Q000494:pharmacology; D012996:Solutions; D013314:Stress, Mechanical; D013395:Sucrose / Q000494:pharmacology
D016054:DNA, Protozoan; D012996:Solutions; D012965:Sodium Chloride; D013395:Sucrose; D019286:Endopeptidase K