An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples

Detalhes bibliográficos
Autor(a) principal: Riediger, Irina Nastassja
Data de Publicação: 2016
Outros Autores: Hoffmaster, Alex R, Casanovas-Massana, Arnau, Biondo, Alexander Welker, Ko, Albert Icksang, Stoddard, Robyn A
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/17792
Resumo: National Institute of Allergy and Infectious Diseases. CAPES (Brazilian Ministry of Education).
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spelling Riediger, Irina NastassjaHoffmaster, Alex RCasanovas-Massana, ArnauBiondo, Alexander WelkerKo, Albert IcksangStoddard, Robyn A2017-02-10T18:23:42Z2017-02-10T18:23:42Z2016RIEDIGER, I. N. et al. An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples. Plos One, v. 11, n. 8, p. e0160523, 2016.1932-6203https://www.arca.fiocruz.br/handle/icict/1779210.1371/journal.pone.0160523National Institute of Allergy and Infectious Diseases. CAPES (Brazilian Ministry of Education).Central Laboratory of the State of Paraná. Curitiba, PR, BrasilNational Center for Emerging and Zoonotic Infectious Diseases. Centers for Disease Control and Prevention, Atlanta, Georgia, USAYale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USAFederal University of the State of Paraná. Department of Veterinary Medicine. Curitiba, PR, BrazilYale University. School of Public Health. Department of Epidemiology of Microbial Diseases. New Haven, Connecticut, USA / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, BA, BrasilNational Center for Emerging and Zoonotic Infectious Diseases. Centers for Disease Control and Prevention, Atlanta, Georgia, USALeptospirosis is a zoonotic disease usually acquired by contact with water contaminated with urine of infected animals. However, few molecular methods have been used to monitor or quantify pathogenic Leptospira in environmental water samples. Here we optimized a DNA extraction method for the quantification of leptospires using a previously described Taqman-based qPCR method targeting lipL32, a gene unique to and highly conserved in pathogenic Leptospira. QIAamp DNA mini, MO BIO PowerWater DNA and PowerSoil DNA Isolation kits were evaluated to extract DNA from sewage, pond, river and ultrapure water samples spiked with leptospires. Performance of each kit varied with sample type. Sample processing methods were further evaluated and optimized using the PowerSoil DNA kit due to its performance on turbid water samples and reproducibility. Centrifugation speeds, water volumes and use of Escherichia coli as a carrier were compared to improve DNA recovery. All matrices showed a strong linearity in a range of concentrations from 106 to 10° leptospires/mL and lower limits of detection ranging from <1 cell /ml for river water to 36 cells/mL for ultrapure water with E. coli as a carrier. In conclusion, we optimized a method to quantify pathogenic Leptospira in environmental waters (river, pond and sewage) which consists of the concentration of 40 mL samples by centrifugation at 15,000×g for 20 minutes at 4°C, followed by DNA extraction with the PowerSoil DNA Isolation kit. Although the method described herein needs to be validated in environmental studies, it potentially provides the opportunity for effective, timely and sensitive assessment of environmental leptospiral burden.engPublic Library of ScienceLeptospiraLeptospiroseMeio ambienteÁguaAnimaisLeptospiraLeptospirosisEnvironmentWaterAnimalsAn Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samplesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/17792/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALRiediger IN An optimized method....pdfRiediger IN An optimized 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dc.title.pt_BR.fl_str_mv An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
title An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
spellingShingle An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
Riediger, Irina Nastassja
Leptospira
Leptospirose
Meio ambiente
Água
Animais
Leptospira
Leptospirosis
Environment
Water
Animals
title_short An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
title_full An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
title_fullStr An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
title_full_unstemmed An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
title_sort An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples
author Riediger, Irina Nastassja
author_facet Riediger, Irina Nastassja
Hoffmaster, Alex R
Casanovas-Massana, Arnau
Biondo, Alexander Welker
Ko, Albert Icksang
Stoddard, Robyn A
author_role author
author2 Hoffmaster, Alex R
Casanovas-Massana, Arnau
Biondo, Alexander Welker
Ko, Albert Icksang
Stoddard, Robyn A
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Riediger, Irina Nastassja
Hoffmaster, Alex R
Casanovas-Massana, Arnau
Biondo, Alexander Welker
Ko, Albert Icksang
Stoddard, Robyn A
dc.subject.other.pt_BR.fl_str_mv Leptospira
Leptospirose
Meio ambiente
Água
Animais
topic Leptospira
Leptospirose
Meio ambiente
Água
Animais
Leptospira
Leptospirosis
Environment
Water
Animals
dc.subject.en.pt_BR.fl_str_mv Leptospira
Leptospirosis
Environment
Water
Animals
description National Institute of Allergy and Infectious Diseases. CAPES (Brazilian Ministry of Education).
publishDate 2016
dc.date.issued.fl_str_mv 2016
dc.date.accessioned.fl_str_mv 2017-02-10T18:23:42Z
dc.date.available.fl_str_mv 2017-02-10T18:23:42Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv RIEDIGER, I. N. et al. An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples. Plos One, v. 11, n. 8, p. e0160523, 2016.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/17792
dc.identifier.issn.pt_BR.fl_str_mv 1932-6203
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pone.0160523
identifier_str_mv RIEDIGER, I. N. et al. An Optimized Method for Quantification of Pathogenic Leptospira in Environmental Water Samples. Plos One, v. 11, n. 8, p. e0160523, 2016.
1932-6203
10.1371/journal.pone.0160523
url https://www.arca.fiocruz.br/handle/icict/17792
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/17792/1/license.txt
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