Comparison of two target genes for detection and genotyping of Giardia lamblia in human feces by PCR and PCR-restriction fragment length polymorphism
Comparison of two target genes for detection and genotyping of Giardia lamblia in human feces by PCR and PCR-restriction fragment length polymorphism
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DOI：
10.1128/jcm.43.12.5940-5944.2005
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发表时间：
2005-12-01
影响因子：
9.4
通讯作者：
Schwartzbrod, J
中科院分区：
文献类型：
--
作者：
Bertrand, I;Albertini, L;Schwartzbrod, J
建立了一种以tpi基因为靶点的PCR检测方法，用于检测人类粪便中的贾第虫并进行基因分型。本试验具有特异性，可区分G. Lamblia组合A和B。G.还用两种先前描述的PCR-限制性片段长度多态性（RFLP）测定法分析了从人粪便中分离的兰氏囊泡，所述测定法基于tpi或gdh基因的检测。这些RFLP分析区分了组合A内的组I和组II或组合B内的组III和组IV。在26例由医院实验室诊断的散发性贾第虫病患者的粪便样本中，我们的PCR检测从25例（96%）中扩增出TPI基因，而当靶向gdh基因时，仅21例（81%）样本呈阳性。在25个阳性样品中，9个（36%）含有组合A，16个（64%）含有组合B。因此，RFLP分析将8个样本（32%）分类为组合A第II组，8个样本（32%）分类为组合B第III组，5个样本（20%）分类为组合B第IV组。无法指定四个样本的组别。tpi和gdh基因在G. Lamblia组合B从15个仅从象牙海岸回来的法国士兵收集的样品中扩增出14个（93%）。所有这些都包含组合B组III。所建立的PCR方法灵敏、简便、特异，表明tpi基因对G. Lamblia基因分型
A PCR assay targeting the tpi gene was developed to detect and to genotype Giardia lamblia in human feces. Our assay was specific and discriminated between G. lamblia assemblages A and B. G. lamblia cysts isolated from human feces were also analyzed with two previously described PCR-restriction fragment length polymorphism (RFLP) assays, which are based on the detection of tpi or gdh genes. These RFLP analyses distinguished groups I and II within assemblage A or groups III and IV within assemblage B. Among 26 fecal samples from patients with sporadic giardiasis diagnosed by hospital laboratories, the tpi gene was amplified from 25 (96%) with our PCR assay, whereas only 21 (81%) samples were positive when the gdh gene was targeted. Of the 25 positive samples, nine (36%) contained assemblage A and 16 (64%) contained assemblage B. Thus, RFLP analysis classified eight samples (32%) in assemblage A group II, eight (32%) in assemblage B group III, and five (20%) in assemblage B group IV. The group could not be specified for four samples. The tpi and gdh genes of G. lamblia assemblage B were amplified from 14 (93%) of 15 samples collected only from French soldiers coming back from the Ivory Coast. All of these contained assemblage B group III. The PCR method developed is sensitive, simple, and specific and shows that the tpi gene is well adapted for G. lamblia genotyping.