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Journal of Clinical Microbiology, July 2008, p. 2252-2262, Vol. 46, No. 7
0095-1137/08/$08.00+0     doi:10.1128/JCM.00116-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Classification of Cryptosporidium Species from Patients with Sporadic Cryptosporidiosis by Use of Sequence-Based Multilocus Analysis following Mutation Scanning{triangledown}

Aaron R. Jex,1* Aradhana Pangasa,1 Bronwyn E. Campbell,1 Margaret Whipp,2 Geoff Hogg,2 Martha I. Sinclair,3 Melita Stevens,4 and Robin B. Gasser1*

Department of Veterinary Science, The University of Melbourne, Werribee, Victoria 3030, Australia,1 Melbourne Diagnostic Unit, Victoria 3052, Australia,2 Department of Epidemiology and Preventive Medicine, Monash University, Central and Eastern Clinical School, Alfred Hospital, Melbourne, Victoria 3004, Australia,3 Melbourne Water Corporation, East Melbourne, Victoria 3002, Australia4

Received 20 January 2008/ Returned for modification 14 April 2008/ Accepted 19 April 2008

In the present study, we analyzed genetic variation in Cryptosporidium species from humans (n = 62) with clinical cryptosporidiosis in South Australia. Sequence variation was assessed in regions within the small subunit of nuclear rRNA (p-SSU), the 70-kDa heat shock protein (p-hsp70), and the 60-kDa glycoprotein (p-gp60) genes by employing single-strand conformation polymorphism analysis and sequencing. Based on the analyses of p-SSU and p-hsp70, Cryptosporidium hominis (n = 38) and Cryptosporidium parvum (n = 24) were identified. The analysis of p-gp60 revealed eight distinct subgenotypes, classified as C. hominis IaA17R1 (n = 3), IbA9G3R2 (n = 14), IbA10G2R2 (n = 20), and IfA12G1R1 (n = 1), as well as C. parvum IIaA18G3R1 (n = 15), IIaA20G3R1 (n = 6), IIaA22G4R1 (n = 2), and IIcA5G3R2 (n = 1). Subgenotypes IaA17R1 and IIaA22G4R1 are new. Of the six other subgenotypes, IbA10G2R2, IIaA18G3R1, IIaA20G3R1, and IIcA5G3R2 were reported previously from the state of Victoria. This is the fourth record in Australia of C. parvum subgenotype IIaA18G3R1 from humans, which, to date, has been isolated only from cattle in other countries. This subgenotype might be a significant contributor to sporadic human cryptosporidiosis and may indicate a greater zoonotic contribution to the infection of humans in the area of study. Comparative analyses revealed, for the first time, the differences in the genetic makeup of Cryptosporidium populations between two relatively close, major metropolitan cities.

* Corresponding author. Mailing address: Department of Veterinary Science, The University of Melbourne, Werribee, Victoria 3030, Australia. Phone: 61 3 9731 2294. Fax: 61 3 9731 2366. E-mail for A. Jex: ajex{at}unimelb.edu.au. E-mail for R. Gasser: robinbg{at}unimelb.edu.au

{triangledown} Published ahead of print on 30 April 2008.

Journal of Clinical Microbiology, July 2008, p. 2252-2262, Vol. 46, No. 7
0095-1137/08/$08.00+0     doi:10.1128/JCM.00116-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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