This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Di Giovanni, G. D.
Right arrow Articles by LeChevallier, M. W.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Di Giovanni, G. D.
Right arrow Articles by LeChevallier, M. W.
Agricola
Right arrow Articles by Di Giovanni, G. D.
Right arrow Articles by LeChevallier, M. W.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, March 2005, p. 1495-1500, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1495-1500.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Quantitative-PCR Assessment of Cryptosporidium parvum Cell Culture Infection

George D. Di Giovanni1* and Mark W. LeChevallier2

American Water Works Service Co., Inc., Belleville, Illinois,1 American Water Works Service Co., Inc., Voorhees, New Jersey2

Received 25 May 2004/ Accepted 12 October 2004

A quantitative TaqMan PCR method was developed for assessing the Cryptosporidium parvum infection of in vitro cultivated human ileocecal adenocarcinoma (HCT-8) cell cultures. This method, termed cell culture quantitative sequence detection (CC-QSD), has numerous applications, several of which are presented. CC-QSD was used to investigate parasite infection in cell culture over time, the effects of oocyst treatment on infectivity and infectivity assessment of different C. parvum isolates. CC-QSD revealed that cell culture infection at 24 and 48 h postinoculation was approximately 20 and 60%, respectively, of the endpoint 72-h postinoculation infection. Evaluation of three different lots of C. parvum Iowa isolate oocysts revealed that the mean infection of 0.1 N HCl-treated oocysts was only 36% of the infection obtained with oocysts treated with acidified Hanks' balanced salt solution containing 1% trypsin. CC-QSD comparison of the C. parvum Iowa and TAMU isolates revealed significantly higher levels of infection for the TAMU isolate, which agrees with and supports previous human, animal, and cell culture studies. CC-QSD has the potential to aid in the optimization of Cryptosporidium cell culture methods and facilitate quantitative evaluation of cell culture infectivity experiments.

* Corresponding author. Present address: Texas A&M University Agricultural Research and Extension Center, El Paso, TX 79927. Phone: (915) 859-9111. Fax: (915) 859-1078. E-mail: gdigiovanni{at}ag.tamu.edu.

Applied and Environmental Microbiology, March 2005, p. 1495-1500, Vol. 71, No. 3
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.3.1495-1500.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Brescia, C. C., Griffin, S. M., Ware, M. W., Varughese, E. A., Egorov, A. I., Villegas, E. N. (2009). Cryptosporidium Propidium Monoazide-PCR, a Molecular Biology-Based Technique for Genotyping of Viable Cryptosporidium Oocysts. Appl. Environ. Microbiol. 75: 6856-6863 [Abstract] [Full Text]  
  • Sifuentes, L. Y., Di Giovanni, G. D. (2007). Aged HCT-8 Cell Monolayers Support Cryptosporidium parvum Infection. Appl. Environ. Microbiol. 73: 7548-7551 [Abstract] [Full Text]  
  • Godiwala, N. T., Vandewalle, A., Ward, H. D., Leav, B. A. (2006). Quantification of In Vitro and In Vivo Cryptosporidium parvum Infection by Using Real-Time PCR.. Appl. Environ. Microbiol. 72: 4484-4488 [Abstract] [Full Text]  
  • Cai, X., Woods, K. M., Upton, S. J., Zhu, G. (2005). Application of Quantitative Real-Time Reverse Transcription-PCR in Assessing Drug Efficacy against the Intracellular Pathogen Cryptosporidium parvum In Vitro. Antimicrob. Agents Chemother. 49: 4437-4442 [Abstract] [Full Text]  
  • Schets, F. M., Engels, G. B., During, M., de Roda Husman, A. M. (2005). Detection of Infectious Cryptosporidium Oocysts by Cell Culture Immunofluorescence Assay: Applicability to Environmental Samples. Appl. Environ. Microbiol. 71: 6793-6798 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»