Plutzer Judit, Karanis Panagiotis, Domokos Klarissza, Törökné Andrea, Márialigeti Károly
National Institute of Environmental Health, Department of Water Hygiene, Gyáli ut 2-6, Budapest H-1096, Hungary.
Int J Hyg Environ Health. 2008 Oct;211(5-6):524-33. doi: 10.1016/j.ijheh.2008.04.004. Epub 2008 Jun 11.
We investigated the prevalence of Giardia and Cryptosporidium species and analysed the genotypes in 36 samples collected from different water sources and various geographic areas in Hungary. Samples were collected from drinking water and sewage treatment plants and from the recreation area of Lake Balaton. The (oo)cysts were purified according to the US EPA 1623 method and they were detected by immunofluorescence test (IFT). Genomic DNA was extracted from all samples and then the GDH target gene for Giardia and the SSUrDNA for both Giardia and for Cryptosporidium species were amplified by PCR. 24 out of 36 samples (67%) were Giardia positive and 15 (42%) were Cryptosporidium positive by IFT. PCR confirmed that 13 out of 36 samples (36%) were Giardia positive and 10 (28%) contained Cryptosporidium. Twelve Giardia and two Cryptosporidium PCR products were successfully sequenced. In seven samples G. lamblia Assemblage A and in one sample Assemblage B and in four cases Assemblages A and B have been found. In one sample C. parvum and in the other separate sample C. meleagridis were detected. Sequence analysis revealed a new subtype of G. duodenalis complex, clustered close to the Assemblage A group. This study provides the first report on simultaneous detection and genotyping of G. duodenalis and Cryptosporidium species from water supplies in Hungary.
我们调查了匈牙利不同水源和不同地理区域采集的36份样本中贾第虫属和隐孢子虫属的流行情况，并分析了其基因型。样本取自饮用水、污水处理厂以及巴拉顿湖的休闲区。（卵）囊根据美国环境保护局1623方法进行纯化，并用免疫荧光试验（IFT）进行检测。从所有样本中提取基因组DNA，然后通过聚合酶链反应（PCR）扩增贾第虫属的谷氨酸脱氢酶（GDH）靶基因以及贾第虫属和隐孢子虫属的小亚基核糖体DNA（SSUrDNA）。IFT检测显示，36份样本中有24份（67%）贾第虫呈阳性，15份（42%）隐孢子虫呈阳性。PCR证实，36份样本中有13份（36%）贾第虫呈阳性，10份（28%）含有隐孢子虫。成功对12份贾第虫和2份隐孢子虫的PCR产物进行了测序。在7份样本中发现了兰氏贾第虫集合体A，1份样本中发现了集合体B，4份样本中同时发现了集合体A和B。在1份样本中检测到微小隐孢子虫，在另一份单独的样本中检测到火鸡隐孢子虫。序列分析揭示了十二指肠贾第虫复合体的一个新亚型，其聚类靠近集合体A组。本研究首次报道了匈牙利供水系统中同时检测十二指肠贾第虫和隐孢子虫属及其基因分型的情况。
