Polymerase chain reaction and nested-PCR approaches for detecting Cryptosporidium in water catchments of water treatment plants in Curitiba, State of Paraná, Brazil
| Autor: | Márcia Benedita Oliveira-Silva, Adriana Oliveira Costa, Juliana Tracz Pereira, Vanete Thomaz Soccol, Silvia Cristina Osaki, Antônio Eduardo Procópio |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Microbiology (medical)
Veterinary medicine lcsh:Arctic medicine. Tropical medicine lcsh:RC955-962 Cryptosporidium Fresh Water DNA Ribosomal Polymerase Chain Reaction law.invention Microbiology Water Purification law Water Supply Nested-PCR parasitic diseases medicine Raw water Polymerase chain reaction biology Sewage Waterborne diseases Water DNA extraction technique Contamination DNA Protozoan medicine.disease biology.organism_classification DNA extraction Infectious Diseases Parasitology Water treatment Nested polymerase chain reaction Brazil |
| Zdroj: | Revista da Sociedade Brasileira de Medicina Tropical v.46 n.3 2013 Revista da Sociedade Brasileira de Medicina Tropical Sociedade Brasileira de Medicina Tropical (SBMT) instacron:SBMT Revista da Sociedade Brasileira de Medicina Tropical, Volume: 46, Issue: 3, Pages: 270-276, Published: JUN 2013 Revista da Sociedade Brasileira de Medicina Tropical, Vol 46, Iss 3, Pp 270-276 (2013) |
| Popis: | Introduction Cryptosporidium is an important protozoan cause of waterborne disease worldwide of concern to public health authorities. To prevent outbreaks of cryptosporidiosis, the monitoring of this parasite in drinking water is necessary. In the present work, the polymerase chain reaction (PCR) and nested-PCR techniques were used to detect Cryptosporidium in raw water from catchment points of four water treatment plants (WTP) in Curitiba, Paraná, Brazil. Methods First, DNA extraction techniques were tested in samples containing decreasing amount of oocysts in reagent water, and PCR and nested-PCR with specific primers for 18SSU rDNA of Cryptosporidium were conducted to determine their sensitivity. In reagent water, a commercial extraction kit provided the best analytical sensitivity, and PCR and nested-PCR allowed the detection of five and two oocysts, respectively, with the primers XIAOR/XIAOF and XIAO1F/XIAO2R. Results In the spiking experiments, only the PCR with the primers AWA995F/AWA1206R was successful at detecting concentrations of 0.1 oocysts/mL. Two catchments samples of raw water and/or water sludge from four WTPs were contaminated with Cryptosporidium. Conclusions The application of the techniques to monitor Cryptosporidium in water and detect contamination in water catchments of WTPs in Curitiba are discussed in the present work. |
| Databáze: | OpenAIRE |
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