title: Giardia intestinalis: DNA extraction approaches to improve PCR results.
abstract: Difficulty in disrupting cysts of Giardia intestinalis, a cosmopolitan protozoan parasite, decreases the yield of DNA extracted and reduces the effectiveness of the polymerase chain reaction (PCR). To improve the detection of the Giardia Glutamate Dehydrogenase (gdh) gene, we re-evaluated the effects of deoxyribonucleic acid (DNA) extraction methods. Purified and concentrated cysts from 33 fecal samples were disrupted using conventional methods, and DNA extraction was conducted using two protocols: the QIAamp Stool Mini Kit and phenol/chloroform/isoamyl alcohol (PCI). PCR amplification was successful for 12 extracted DNA samples (36%) using PCI following a glass bead and freeze/thaw pretreatment and for all 33 samples (100%) using the QIAamp Stool Mini Kit following the aforementioned pretreatment. Consequently, the pretreatment of cysts with glass beads and freeze/thaw cycles followed by extraction of DNA with the QIAamp Stool Mini kit was the more effective protocol.
authors.affiliation: Department of Parasitology and Mycology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran.
authors.forename: Zahra
authors.initials: Z
authors.lastname: Babaei
authors.forename: Hormozd
authors.initials: H
authors.lastname: Oormazdi
authors.forename: Sasan
authors.initials: S
authors.lastname: Rezaie
authors.forename: Mostafa
authors.initials: M
authors.lastname: Rezaeian
authors.forename: Elham
authors.initials: E
authors.lastname: Razmjou
mesh_terms: D016054:DNA, Protozoan / Q000737:chemistry / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005243:Feces / Q000469:parasitology*; D005838:Genotype; D016829:Giardia lamblia / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification; D005873:Giardiasis / Q000469:parasitology*; D005969:Glutamate Dehydrogenase / Q000235:genetics; D006801:Humans; D016133:Polymerase Chain Reaction / Q000592:standards*
chemical_list: D016054:DNA, Protozoan; D005969:Glutamate Dehydrogenase