ArticleEcotoxicology (London, England)2009
Development of a direct DNA extraction protocol for real-time PCR detection of Giardia lamblia from surface water.
Article in Ecotoxicology (London, England), 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed, 24 citations in OpenAlex.
- Safe drinking water: to what extent are shallow wells reliable?Journal of water and health · 2024Article
- Occurrence of Giardia duodenalis in selected stations and tributary rivers of Laguna Lake, Philippines.Environmental monitoring and assessment · 2021Article
- Detection and molecular characterization of Cryptosporidium species and Giardia assemblages in two watersheds in the metropolitan region of São Paulo, Brazil.Environmental science and pollution research international · 2018Article
- Presence of Cryptosporidium parvum and Giardia lamblia in water samples from Southeast Asia: towards an integrated water detection system.Infectious diseases of poverty · 2016Article
- Spatial variations of bacterial community and its relationship with water chemistry in Sanya Bay, South China Sea as determined by DGGE fingerprinting and multivariate analysis.Ecotoxicology (London, England) · 2015Article
- Spatial variation of bacterial community structure of the Northern South China Sea in relation to water chemistry.Ecotoxicology (London, England) · 2012Article
- Genotyping of Giardia duodenalis cysts by new real-time PCR assays for detection of mixed infections in human samples.Applied and environmental microbiology · 2010Article
- A Simple and Cost-Effective Freeze-Thaw Based Method forIranian journal of parasitologyArticle
Corrections and comments
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Authors and funding
4 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Giardia lamblia is one of the most recognized waterborne protozoan parasites causing gastrointestinal disease. A simple but effective DNA extraction protocol for real-time PCR detection from surface water samples was developed in this study. Eleven protocols were compared, which consisted of freeze-thaw treatments (liquid N(2) and boiling water) and purification using the Qiagen DNeasy kit, together with different combinations of proteinase K, PVP360, GITC and Chelex 100 incubation. Using concentrated surface water samples spiked with G. lamblia cysts, the necessary steps for high DNA recovery were shown to be freeze-thaw, DNeasy purification and Chelex 100 incubation. Multiple rounds of freeze-thaw treatment (five cycles per round) were reported for the first time in this study to significantly increase the DNA yield from G. lamblia cysts, from ~20% after one round of freeze-thaw to 40 and 70% after two and three-rounds of freeze-thaw, respectively. More than three rounds of freeze-thaw treatment did not promote additional DNA recovery. The final protocol included three-three-rounds of freeze-thaw treatment, DNeasy purification and Chelex 100 incubation. This method was simpler, more cost-effective, and had a comparable DNA recovery to methods involving immunomagnetic separation.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.