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Journal of Clinical Microbiology, March 2003, p. 981-992, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.981-992.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Epidemiological Analysis of Cryptosporidium Isolates from Humans and Animals by Using a Heteroduplex Mobility Assay and Nucleic Acid Sequencing Based on a Small Double-Stranded RNA Element

Francesca Leoni,1,2,3 Chris I. Gallimore,4 Jonathan Green,4,{dagger} and Jim McLauchlin1*

PHLS Food Safety Microbiology Laboratory,1 PHLS Enteric, Respiratory, and Neurological Virus Laboratory, Central Public Health Laboratory, London NW9 5HT, United Kingdom,4 School of Chemical and Life Sciences, University of Greenwich, Woolwich, London SE18 8PF, United Kingdom,2 IZS-UM Sezione Diagnostica di Fermo, Istituto Zooprofilattico Sperimentale dell'Umbria e delle Marche, Fermo 63023, Italy3

Received 9 September 2002/ Returned for modification 17 October 2002/ Accepted 9 December 2002

Two extrachromosomal double-stranded RNA (dsRNA) elements occur in Cryptosporidium parvum. A heteroduplex mobility assay (HMA) was developed for the rapid characterization of sequence diversity in a 173-bp fragment of the small dsRNA element of Cryptosporidium with either a natural sequence from Cryptosporidium meleagridis or a synthetic sequence as reference DNA. The 173-bp fragment was generated from 265 samples of whole feces (242 from humans and 18 from livestock with C. parvum genotype 1 or 2, 4 from humans with Cryptosporidium felis, and 1 from a human with C. meleagridis). The HMA method identified 21 patterns in C. parvum (8 in genotype 1, 12 in genotype 2, and a type common to both genotypes), 4 patterns in C. felis, and 1 pattern in C. meleagridis. All patterns were confirmed as distinct by DNA sequencing. For genotype 1, a single HMA type was found in 89% of samples: 64 of 65 cases from three waterborne outbreaks, all 16 cases from eight intrafamilial outbreaks, and 17 of 28 sporadic cases. Among the remaining 11 sporadic cases due to genotype 1, seven other HMA types were detected. For genotype 2, a single HMA type was found in 72% of samples: 36 of 43 cases from three waterborne outbreaks, 11 of 15 cases from seven intrafamilial outbreaks, 44 of 75 sporadic cases, and all 18 samples from livestock. Within the intrafamilial outbreaks, two other HMA types were identified: the same HMA type was detected in samples from cases within the same outbreak. Among the sporadic cases due to genotype 2, 10 additional HMA types were detected.

* Corresponding author. Mailing address: PHLS Food Safety Microbiology Laboratory, Central Public Health Laboratory, 61 Colindale Ave., London NW9 5HT, United Kingdom. Phone: 44-208-200-4400. Fax: 44-208-2358-3112. E-mail: jmclauchlin{at}phls.org.uk.

{dagger} Present address PHLS Bioinformatics Unit, Central Public Health Laboratory, London NW9 5HT, United Kingdom.

Journal of Clinical Microbiology, March 2003, p. 981-992, Vol. 41, No. 3
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.3.981-992.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Leoni, F., Mallon, M. E., Smith, H. V., Tait, A., McLauchlin, J. (2007). Multilocus Analysis of Cryptosporidium hominis and Cryptosporidium parvum Isolates from Sporadic and Outbreak-Related Human Cases and C. parvum Isolates from Sporadic Livestock Cases in the United Kingdom. J. Clin. Microbiol. 45: 3286-3294 [Abstract] [Full Text]  
  • Learmonth, J. J., Ionas, G., Ebbett, K. A., Kwan, E. S. (2004). Genetic Characterization and Transmission Cycles of Cryptosporidium Species Isolated from Humans in New Zealand. Appl. Environ. Microbiol. 70: 3973-3978 [Abstract] [Full Text]  


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