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Appl Environ Microbiol, April 1998, p. 1454-1458, Vol. 64, No. 4
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Failure To Differentiate Cryptosporidium parvum from C. meleagridis Based on PCR Amplification of Eight DNA Sequences

Dominique Champliaud,1,2 Philippe Gobet,1 Muriel Naciri,3 Odile Vagner,1 José Lopez,1,2 Jean Christophe Buisson,1 Istvan Varga,4 Géraldine Harly,5 Roseline Mancassola,3 and Alain Bonnin1,2,*

Laboratoire de Parasitologie et Mycologie, Hôpital du Bocage,1 and Laboratoire de Microbiologie Médicale et Moléculaire, Université de Bourgogne,2 21034 Dijon Cedex, INRA de Tours, 37380 Nouzilly,3 and Laboratoire Départemental de la Côte d'Or, Dijon,5 France and University of Veterinary Sciences, Budapest, Hungary4

Received 6 October 1997/Accepted 15 January 1998

In order to determine the specificities of PCR-based assays used for detecting Cryptosporidium parvum DNA, eight pairs of previously described PCR primers targeting six distinct regions of the Cryptosporidium genome were evaluated for the detection of C. parvum, the agent of human cryptosporidiosis, and C. muris, C. baileyi, and C. meleagridis, three Cryptosporidium species that infect birds or mammals but are not considered to be human pathogens. The four Cryptosporidium species were divided into two groups: C. parvum and C. meleagridis, which gave the same-sized fragments with all the reactions, and C. muris and C. baileyi, which gave positive results with primer pairs targeting the 18S rRNA gene only. In addition to being genetically similar at each of the eight loci analyzed by DNA amplification, C. parvum and C. meleagridis couldn't be differentiated even after restriction enzyme digestion of the PCR products obtained from three of the target genes. This study indicates that caution should be exercised in the interpretation of data from water sample analysis performed by these methods, since a positive result does not necessarily reflect a contamination by the human pathogen C. parvum.

* Corresponding author. Mailing address: Laboratoire de Parasitologie et Mycologie, Hôpital du Bocage, 21034 Dijon Cedex, France. Phone: 33 (0)3 80 29 36 03. Fax: 33 (0)3 80 29 32 80. E-mail: APBonnin{at}aol.com.




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