Literature DB >> 27095569

An easy 'one tube' method to estimate viability of Cryptosporidium oocysts using real-time qPCR.

A Paziewska-Harris1, G Schoone2, H D F H Schallig2.   

Abstract

Viability estimation of the highly resistant oocysts of Cryptosporidium remains a key issue for the monitoring and control of this pathogen. We present here a simple 'one tube' quantitative PCR (qPCR) protocol for viability estimation using a DNA extraction protocol which preferentially solubilizes excysted sporozoites rather than oocysts. Parasite DNA released from excysted sporozoites was quantified by real-time qPCR using a ribosomal DNA marker. The qPCR signal was directly proportional to the number of oocysts excysted, and a power-law relationship was noted between oocyst age and the proportion excysting. Unexcysted oocysts released negligible amounts of DNA making the method suitable for estimating viability of as few as 10 oocysts.

Keywords:  Cryptosporidium parvum; Environmental samples; Excystation; Sporozoites; Water quality monitoring

Mesh：

Substances：

Year:  2016        PMID： 27095569     DOI： 10.1007/s00436-016-5044-z

Source DB:  PubMed          Journal:  Parasitol Res        ISSN： 0932-0113            Impact factor:   2.289


  19 in total

1.  Rapid and simple method for purification of nucleic acids.

Authors:  R Boom; C J Sol; M M Salimans; C L Jansen; P M Wertheim-van Dillen; J van der Noordaa
Journal:  J Clin Microbiol       Date:  1990-03       Impact factor: 5.948

Review 2.  Critical processes affecting Cryptosporidium oocyst survival in the environment.

Authors:  B J King; P T Monis
Journal:  Parasitology       Date:  2006-11-13       Impact factor: 3.234

3.  Quantitative analysis of Cryptosporidium growth in in vitro culture--the impact of parasite density on the success of infection.

Authors:  Anna Paziewska-Harris; Martin Singer; Gerard Schoone; Henk Schallig
Journal:  Parasitol Res       Date:  2015-10-05       Impact factor: 2.289

4.  Number of Cryptosporidium parvum oocysts or Giardia spp cysts shed by dairy calves after natural infection.

Authors:  D V Nydam; S E Wade; S L Schaaf; H O Mohammed
Journal:  Am J Vet Res       Date:  2001-10       Impact factor: 1.156

5.  Quantification of Cryptosporidium parvum in anaerobic digesters treating manure by (reverse-transcription) quantitative real-time PCR, infectivity and excystation tests.

Authors:  G Garcés; M Effenberger; M Najdrowski; C Wackwitz; A Gronauer; P A Wilderer; M Lebuhn
Journal:  Water Sci Technol       Date:  2006       Impact factor: 1.915

6.  Composition and conformation of Cryptosporidium parvum oocyst wall surface macromolecules and their effect on adhesion kinetics of oocysts on quartz surface.

Authors:  Yuanyuan Liu; Mark S Kuhlenschmidt; Theresa B Kuhlenschmidt; Thanh H Nguyen
Journal:  Biomacromolecules       Date:  2010-08-09       Impact factor: 6.988

7.  Cryptosporidium hominis: experimental challenge of healthy adults.

Authors:  Cynthia L Chappell; Pablo C Okhuysen; Rebecca Langer-Curry; Giovanni Widmer; Donna E Akiyoshi; Sultan Tanriverdi; Saul Tzipori
Journal:  Am J Trop Med Hyg       Date:  2006-11       Impact factor: 2.345

8.  Combination of cell culture and quantitative PCR for screening of drugs against Cryptosporidium parvum.

Authors:  M Shahiduzzaman; V Dyachenko; A Obwaller; S Unglaube; A Daugschies
Journal:  Vet Parasitol       Date:  2009-03-13       Impact factor: 2.738

9.  Real-time PCR for quantification of Giardia and Cryptosporidium in environmental water samples and sewage.

Authors:  Rebecca A Guy; Pierre Payment; Ulrich J Krull; Paul A Horgen
Journal:  Appl Environ Microbiol       Date:  2003-09       Impact factor: 4.792

10.  Cryptosporidium parvum: determination of ID₅₀ and the dose-response relationship in experimentally challenged dairy calves.

Authors:  J A Zambriski; D V Nydam; Z J Wilcox; D D Bowman; H O Mohammed; J L Liotta
Journal:  Vet Parasitol       Date:  2013-04-20       Impact factor: 2.738
View more
  2 in total

Review 1.  Assessing viability and infectivity of foodborne and waterborne stages (cysts/oocysts) of Giardia duodenalis, Cryptosporidium spp., and Toxoplasma gondii: a review of methods.

Authors:  Angélique Rousseau; Stéphanie La Carbona; Aurélien Dumètre; Lucy J Robertson; Gilles Gargala; Sandie Escotte-Binet; Loïc Favennec; Isabelle Villena; Cédric Gérard; Dominique Aubert
Journal:  Parasite       Date:  2018-03-19       Impact factor: 3.000

Review 2.  Transmission of waterborne parasites in the Association of Southeast Asian Nations (ASEAN): Overview and direction forward.

Authors:  Yvonne A L Lim; Veeranoot Nissapatorn
Journal:  Food Waterborne Parasitol       Date:  2017-08-12
  2 in total

北京卡尤迪生物科技股份有限公司 © 2022-2023.