title: Recovery of DNA of Giardia intestinalis cysts from surface water concentrates measured with PCR and real time PCR.
abstract: The most important restriction for the detection in water samples is the low concentration of Giardia intestinalis cysts, additional difficulty is the presence of PCR inhibitors. We have carried out trials in order to assess the sensitivity of semi-nested PCR and TaqMan real time PCR on the basis of DNA extracted from G. intestinalis cysts coming from spiked environmental and distilled water samples, filtrated with the use of Filta-Max® equipment (1623 Method). Removal of inhibitors was carried out with addition of BSA in different concentrations. During the filtration and concentration of water samples, losses of cysts have been recorded. Moreover, addition of BSA to the PCR and real time PCR mix increases the sensitivity of reaction. The optimal concentration of BSA for semi‑nested PCR was 15 and 20 ng/μl, whereas for real time PCR 5 ng/μl.
authors.affiliation: Department of Genetics, University of Szczecin, ul. Felczaka 3c, 71-412 Szczecin, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Adamska
authors.forename: A
authors.initials: A
authors.lastname: Leońska-Duniec
authors.forename: A
authors.initials: A
authors.lastname: Maciejewska
authors.forename: M
authors.initials: M
authors.lastname: Sawczuk
authors.forename: B
authors.initials: B
authors.lastname: Skotarczak
mesh_terms: D016054:DNA, Protozoan / Q000302:isolation & purification*; D005618:Fresh Water / Q000469:parasitology*; D016829:Giardia lamblia / Q000235:genetics* / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000592:standards*; D060888:Real-Time Polymerase Chain Reaction / Q000592:standards*; D012680:Sensitivity and Specificity
chemical_list: D016054:DNA, Protozoan
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