Detection and characterisation of Giardia and Cryptosporidium in Hungarian raw, surface and sewage water samples by IFT, PCR and sequence analysis of the SSUrRNA and GDH genes

Plutzer, Judit; Karanis, Panagiotis; Domokos, K.; Törökné, Andrea; Màrialigeti, Károly

doi:10.1016/j.ijheh.2008.04.004

Cited by 77 publications

(55 citation statements)

References 31 publications

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“…However, nine samples were microscopically positive but negative by PCR in this study. Similar results were also obtained in some other studies conducted in Hungary (Plutzer et al 2008), Spain (Castro-Hermida et al 2008, France (Coupe et al 2006), and Finland (Hanninen et al 2005). This low ratio of positive PCR result may be related to the persistence of potent inhibitor or other interfering compounds in the DNA eluate or to the heterogenous distribution of low oocyst numbers in the samples.…”

Section: Initial Precision and Recovery Of Cryptosporidium Parvum Oocsupporting

confidence: 87%

Occurrences and genotypes of Cryptosporidium oocysts in river network of southern-eastern China

Xiao

Chen

et al. 2011

Parasitol Res

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Transportation of Cryptosporidium oocysts in river type source water is of great concern in an area where extensive human activities exist. In this study, a total of 47 samples were collected from Tongxiang, China, where drinking source water was taken from a complicated river network system, by three sampling campaigns over a rainy season in 2009, to reveal the presence, genotypes, and likely source of Cryptosporidium oocysts in river water. Immunofluorescence microscopy analyses show that 37 (78.7%) were Cryptosporidium positive, with a mean concentration of 0.51 oocysts per liter. These results suggest that the protozoa were commonly distributed in the river network type source water of Tongxiang with a relatively low concentration level. PCR analysis was used to determine the species/genotypes of Cryptosporidium, which revealed the presence of the animal related species/ genotypes including Cryptosporidium suis, Cryptosporidium fragile, and the avian III, pig II, cervine genotypes. Three of them were also detected in wastewater samples taken from neighboring animal farms, showing that farm animals rather than human might be the major pollution sources. This is the first report on simultaneous detection and genotyping of Cryptosporidium oocysts from surface water in China.

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Section: Initial Precision and Recovery Of Cryptosporidium Parvum Oocsupporting

confidence: 87%

Occurrences and genotypes of Cryptosporidium oocysts in river network of southern-eastern China

Xiao

Chen

et al. 2011

Parasitol Res

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“…Extraction of Genomic DNA and Molecular Detection by PCR and LAMP Total DNA, representing 50% of the sucrose purified water concentrates, was extracted by the QIAamp DNA Mini Kit (Qiagen GmbH, Hilden, Germany) according to the modified protocol of Plutzer et al (2008) with additional processing of 10 freeze-thaw cycles after the resuspension step in lysis buffer, supplied from the manufacturer. All samples were frozen by immersion in liquid nitrogen for 1 minute and thawed in a water bath with boiling water, until the frozen samples were completely dissolved.…”

Section: Microscopic Detection and Identification Of Cryptosporidium mentioning

confidence: 99%

“…The nested PCR targeting the small subunit (SSU) rRNA gene was performed as described by Xiao et al (1999;2001) and Plutzer et al (2008). Both PCRs were performed in standard mixtures of 50 ml containing 200 nmoles of each primer, 200 mM dNTP of each dNTP (Finnzymes, Espoo, Finland), 16 PCR buffer containing 1.5 mM MgCl 2 (Qiagen GmbH), 3 mM MgCl 2 (Qiagen GmbH), 2.5 U HotstarTaq DNA polymerase (Qiagen GmbH) and 2 ml bovine serum albumin (acetylated, 10 mg/ ml) (Promega, Madison, WI, USA).…”

Section: Microscopic Detection and Identification Of Cryptosporidium mentioning

confidence: 99%

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

Kolören

Sotiriadou

Karanis

2011

Annals of Tropical Medicine & Parasitology

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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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“…Genomic DNA was isolated by the QIAamp DNA Mini Kit (Qiagen) as described by the manufacturer, following the addition of 10 freeze-thaw cycles according to Plutzer et al [9]. All samples were frozen by immersion in liquid nitrogen for 1 min and thawed in a water bath with boiling water, until the frozen samples were completely dissolved.…”

Section: Genomic Dna Extractionmentioning

confidence: 99%

LAMP DETECTION of Cryptosporidium SPECIES in SURFACE WATER SAMPLES COLLECTED from RIVER YESILIRMAK and STREAM TERSAKAN (Samsun-Amasya)

Kolören¹

2017

ANADOLU UNIVERSITY JOURNAL OF SCIENCE AND TECHNOLOGY –C Life Sciences and Biotechnology

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Detection and characterisation of Giardia and Cryptosporidium in Hungarian raw, surface and sewage water samples by IFT, PCR and sequence analysis of the SSUrRNA and GDH genes

Cited by 77 publications

References 31 publications

Occurrences and genotypes of Cryptosporidium oocysts in river network of southern-eastern China

Occurrences and genotypes of Cryptosporidium oocysts in river network of southern-eastern China

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

LAMP DETECTION of Cryptosporidium SPECIES in SURFACE WATER SAMPLES COLLECTED from RIVER YESILIRMAK and STREAM TERSAKAN (Samsun-Amasya)

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