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 Ochiai, Y.
Right arrow Articles by Hosaka, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ochiai, Y.
Right arrow Articles by Hosaka, M.
Agricola
Right arrow Articles by Ochiai, Y.
Right arrow Articles by Hosaka, M.

 Previous Article  |  Next Article 

Applied and Environmental Microbiology, February 2005, p. 898-903, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.898-903.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Detection and Discrimination of Cryptosporidium parvum and C. hominis in Water Samples by Immunomagnetic Separation-PCR

Yoshitsugu Ochiai,1* Chieko Takada,1 and Mitsugu Hosaka1

Division of Water Environment, Tokyo Metropolitan Institute of Public Health, Shinjuku-ku, Tokyo, Japan1

Received 16 May 2004/ Accepted 21 September 2004

Cryptosporidium parvum and C. hominis have been the cause of large and serious outbreaks of waterborne cryptosporidiosis. A specific and sensitive recovery-detection method is required for control of this pathogen in drinking water. In the present study, nested PCR-restriction fragment length polymorphism (RFLP), which targets the divergent Cpgp40/15 gene, was developed. This nested PCR detected only the gene derived from C. parvum and C. hominis strains, and RFLP was able to discriminate between the PCR products from C. parvum and C. hominis. To evaluate the sensitivity of nested PCR, C. parvum oocysts inoculated in water samples of two different turbidities were recovered by immunomagnetic separation (IMS) and detected by nested PCR and fluorescent antibody assay (FA). Genetic detection by nested PCR and oocyst number confirmed by FA were compared, and the results suggested that detection by nested PCR depends on the confirmed oocyst number and that nested PCR in combination with IMS has the ability to detect a single oocyst in a water sample. We applied an agitation procedure with river water solids to which oocysts were added to evaluate the recovery and detection by the procedure in environmental samples and found some decrease in the rate of detection by IMS.

* Corresponding author. Present address: Department of Veterinary Public Health, Nippon Veterinary and Animal Science University, 1-7-1 Kyonan, Musashino, Tokyo 180-8602, Japan. Phone: 81-422-31-4151, ext. 282. Fax: 81-422-30-7531. E-mail: yochiai{at}nvau.ac.jp.

Applied and Environmental Microbiology, February 2005, p. 898-903, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.898-903.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Frazar, C. D., Orlandi, P. A. (2007). Evaluation of Two DNA Template Preparation Methods for Post-Immunomagnetic Separation Detection of Cryptosporidium parvum in Foods and Beverages by PCR. Appl. Environ. Microbiol. 73: 7474-7476 [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»