Investigations and comparative detection of Cryptosporidium species by microscopy, nested PCR and LAMP in water supplies of Ordu, Middle Black Sea, Turkey.

Z Koloren, I Sotiriadou, P Karanis
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引用次数: 35

Abstract

A total of 70 water samples, including tap, river, fountain and well water were collected in the Ordu province, Middle Black Sea, Turkey and investigated for the detection of Cryptosporidium oocysts. The samples were directly screened microscopically for Cryptosporidium oocysts' detection by immunofluorescence test and subsequently DNA was extracted for the molecular detection by loop-mediated isothermal amplification (LAMP) and nested polymerase chain reaction (PCR). Eighteen out of the 70 (25·7%) water samples were found positive for Cryptosporidium spp. by immunofluorescence test and 19 (27·1%) were found positive by LAMP. Nested PCR products were not generated in any of the investigated water samples. A total of 16 randomly selected pellets were spiked with 10 Cryptosporidium oocysts to test the efficiency of the applied method. All the samples were found positive by LAMP for the presence of Cryptosporidium DNA, while the nested PCR assay was positive in only seven (43·75%) out of the 16 examined spiked samples. This is the first report on the occurrence of Cryptosporidium species in environmental and drinking water supplies in the Black Sea area.

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镜检法、巢式PCR法和LAMP法对土耳其中黑海Ordu地区水源隐孢子虫种类的调查及比较检测。
在土耳其黑海中部Ordu省采集自来水、河流、喷泉、井水等70份水样,对隐孢子虫卵囊进行检测。直接镜检隐孢子虫卵囊,采用免疫荧光检测,提取DNA,采用环介导等温扩增(LAMP)和巢式聚合酶链反应(PCR)进行分子检测。70份水样中隐孢子虫免疫荧光检测阳性18份(25.7%),LAMP检测阳性19份(27.1%)。在任何调查的水样中都没有产生巢式PCR产物。随机抽取16粒微球,加入10个隐孢子虫卵囊,检验该方法的有效性。所有样品的LAMP检测结果均为隐孢子虫DNA阳性,而巢式PCR检测结果仅为7份(43.75%)。本文首次报道了黑海地区环境和饮用水中隐孢子虫的发生情况。
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Annals of tropical medicine and parasitology
Annals of tropical medicine and parasitology 医学-公共卫生、环境卫生与职业卫生
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