Giardia intestinalis: DNA extraction approaches to improve PCR results

被引：43
作者：
Babaei, Zahra [1 ,3 ]
Oormazdi, Hormozd [1 ]
Rezaie, Sasan [2 ]
Rezaeian, Mostafa [2 ]
Razmjou, Elham [1 ]
机构：
[1] Univ Tehran Med Sci, Sch Med, Dept Parasitol & Mycol, Tehran, Iran
[2] Univ Tehran Med Sci, Sch Publ Hlth, Dept Med Parasitol & Mycol, Tehran, Iran
[3] Kerman Univ Med Sci, Sch Med, Dept Parasitol, Kerman, Iran
来源：
EXPERIMENTAL PARASITOLOGY | 2011年 / 128卷 / 02期
关键词：
Flagellate protozoa; DNA extraction method; Glass beads; PCR; Glutamate Dehydrogenase gene; REAL-TIME PCR; GLUTAMATE-DEHYDROGENASE GENE; FRAGMENT LENGTH POLYMORPHISM; STOOL SPECIMENS; FECAL SAMPLES; HUMAN FECES; ENTAMOEBA-HISTOLYTICA; LAMBLIA; DUODENALIS; ASSAY;
D O I：
10.1016/j.exppara.2011.02.001
中图分类号：
R38 [医学寄生虫学]; Q [生物科学];
学科分类号：
07 ; 0710 ; 09 ; 100103 ;
摘要：
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. (C) 2011 Elsevier Inc. All rights reserved.
引用
收藏
页码：159 / 162
页数：4
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