A multi-locus study of cryptosporidium parasites isolated from patients living in iran, Malawi, Nigeria, the United kingdom, and Vietnam.

  • Salman Ghaffari Dept. of Parasitology and Mycology, School of Medicine, Babol University of Medical Sciences, Babol, Iran.
  • Narges Kalantari Cellular and Molecular Biology Research Centre, Babol University of Medical , Babol, Iran.
Keywords: Cryptosporidium, Genotyping, Multilocus study, Subgenotyping


 Background: Cryptosporidium species are important cause of diarrheal diseases in both developing and developed countries. This study aimed to compare the perfor-mance of several molecular methods for identification of Cryptosporidium species, and to detect genetic variation among each of these species isolated from Iran, Ma-lawi, Nigeria, Vietnam and the United Kingdom. Methods: The oocysts DNA samples were derived from 106 Cryptosporidium posi-tive feces. Polymerase chain reaction, PCR- restriction fragment length polymor-phism and DNA sequence analysis of the 18S rRNA and the Cryptosporidium oo-cysts wall protein genes; PCR and DNA sequence analysis of a fragment of 70 kDa heat shock protein and 60 kDa glycoprotein genes were carried out. Results: Based on these analysis, three species of Cryptosporidium including C. homi-nis, C. parvum and C. meleagridis, and both C. hominis and C. parvum were found in Iranian and the UK samples, respectively. Also, three C. hominis (Ib, Ib3& Id) and three C. parvum (IIa, IIc & IId) subtypes were identified by sequence analysis of the GP60 gene. Of these, C. hominis Ib was predominant and interestingly, one subgen-otype (C. hominis Ib A10G2) accounted for the majority of the samples. Conclusion: The current study demonstrates the complex subtypes of Cryptosporid-ium isolates in both developing and developed countries. This is the first report of C. parvum IId subgenotype and three new subtypes of C. parvum IIa in the UK, a new subtype of C. hominis Id from Malawi; and the first multi-locus study of three species of Cryptosporidium in human from Iran.


Xiao L. Molecular epidemiology of crypto-sporidiosis: an update. Exp Parasitol. 2010; 124: 80-89.

Xiao L, Ryan UM. Cryptosporidiosis: an update in molecular epidemiology. Curr Opin Infect Dis. 2004; 17: 483-490.

Cacciò SM. Molecular epidemiology of human cryptosporidiosis. Parassitologia. 2005; 47: 185-192.

Xiao L, Feng Y. Zoonotic cryptosporidiosis. FEMS Immunol Med Microbiol. 2008; 52: 309-323.

Xiao L, Morgan UM, Limor J, et al. Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species. Appl Environ Microbiol. 1999; 65: 3386-3391.

Xiao L, Bern C, Limor J, et al. Identification of 5 types of Cryptosporidium parasites in children in Lima, Peru. J Infect Dis. 2001; 183: 492-497.

Spano F, Putignani L, McLauchlin J, Casemore DP and Crisanti A. PCR‐RFLP analysis of the Cryptosporidium oocyst wall protein (COWP) gene discriminates between C. wrairi and C. parvum, and between C. parvum isolates of human and animal origin. FEMS Microbiol Lett. 1997; 150: 209-217.

Gatei W, Greensill J, Ashford RW, et al. Molecular analysis of the 18S rRNA gene of Cryptosporidium parasites from patients with or without human immunodeficiency virus infections living in Kenya, Malawi, Brazil, the United Kingdom, and Vietnam. J Clin Microbiol. 2003; 41: 1458-1462.

Meamar AR, Guyot K, Certad G, et al. Molecular characterization of Cryptosporidium isolates from humans and animals in Iran. Appl Environ Microbiol. 2007; 73: 1033-1035.

Chalmers R, Robinson G, Elwin K, et al. Detection of Cryptosporidium species and sources of contamination with Cryptosporidium hominis during a waterborne outbreak in north west Wales. J Water Health. 2010; 8: 311-325.

Zintl A, Proctor AF, Read, C et al. The prevalence of Cryptosporidium species and subtypes in human faecal samples in Ireland. Epidemiol Infect. 2009; 137: 270.

Gatei W, Das P, Dutta P, et al. Multilocus sequence typing and genetic structure of Cryptosporidium hominis from children in Kolkata, India. Infect Genet Evol. 2007; 7: 197-205.

Sulaiman IM, Lal AA, Xiao L. A population genetic study of the Cryptosporidium parvum human genotype parasites. J Eukaryot Microbiol. 2001; 48: 24s-27s.

Gatei W, Wamae CN, Mbae C, et al. Cryptosporidiosis: prevalence, genotype analy-sis, and symptoms associated with infections in children in Kenya. Am J Trop Med Hyg. 2006; 75: 78-82.

Peng MM, Matos O, Gatei W, et al. A comparison of Cryptosporidium subgenotypes from several geographic regions. J Eukaryot Microbiol. 2001; 48: 28s-31s.

Leav BA, Mackay MR, Anyanwu A, et al. Analysis of sequence diversity at the highly polymorphic Cpgp40/15 locus among Cryptosporidium isolates from human immuno-deficiency virus-infected children in South Africa. Infect Immun. 2002; 70: 3881-3890.

Akiyoshi DE, Tumwine JK, Bakeera-Kitaka S, Tzipori S. Subtype analysis of Cryptosporidium isolates from children in Uganda. J Parasitol. 2006; 92: 1097-1100.

Peng MM, Meshnick SR, Cunliffe NA, et al. Molecular epidemiology of cryptosporidiosis in children in Malawi. J Eukaryot Microbiol. 2003; 50: 557-559.

Cama VA, Ross JM, Crawford S, et al. Differences in clinical manifestations among Cryptosporidium species and subtypes in HIV-infected persons. J Infect Dis. 2007; 196: 684-691.

Ghaffari S, Kalantari N. Molecular analysis of 18S rRNA gene of Cryptosporidium parasites from patients living in Iran, Malawi, Nigeria and Vietnam. Int J Mol Cell Med.2012; 1: 153-161

Zhou L, Singh A, Jiang J, Xiao L. Molecular surveillance of Cryptosporidium spp. in raw wastewater in Milwaukee: implications for understanding outbreak occurrence and trans-mission dynamics. J Clin Microbiol. 2003; 41: 5254-5257.

Dereeper A, Guignon V, Blanc G, et al. Phy-logeny.fr: robust phylogenetic analysis for the non-specialist. Nucleic Acids Res. 2008; 1, 36 (Web Server Issue):W465-9.

Monteiro L, Bonnemaison D, Vekris A, et al. Complex polysaccharides as PCR inhibitors in feces :Helicobacter pylori model. J Clin Microbiol 1997;35:995-8.

Chalmers RM, Smith R, Elwin K, Clifton-Hadley FA, Giles M. Epidemiology of anthroponotic and zoonotic human cryptosp-oridiosis in England and Wales, 2004–2006. Epidemiol Infect. 2011; 139: 700-712.

Chalmers RM, Elwin K, Thomas AL, Joynson DH. Infection with unusual types of Cryptosporidium is not restricted to immunoc-ompromised patients. J Infect Dis. 2002; 185: 270-271.

Leoni F, Amar C, Nichols G, Pedraza-Diaz S, McLauchlin J. Genetic analysis of Cry-ptosporidium from 2414 humans with diarrhoea in England between 1985 and 2000. J Med Microbiol. 2006; 55: 703-707.

Chalmers RM, Ferguson C, Cacciò S, et al. Direct comparison of selected methods for genetic categorisation of Cryptosporidium parvum and Cryptosporidium hominis species. Int J Parasitol. 2005; 35: 397-410.

Sulaiman IM, Hira PR, Zhou L, et al. Unique endemicity of cryptosporidiosis in children in Kuwait. J Clin Microbiol. 2005; 43: 2805-2809.

Abu Samra N, Thompson PN, Jori F, et al. Genetic characterization of Cryptosporidium spp. in diarrhoeic children from four provinces in South Africa. Zoonoses Public Health. 2013; 60: 154-159.

Taghipour N, Nazemalhosseini-Mojarad E, Haghighi A, et al. Molecular epidemiology of cryptosporidiosis in Iranian children, Tehran, Iran. Iran J Parasitol. 2011; 6: 41-45.

Sulaiman IM, Morgan UM, Thompson RC, Lal AA, Xiao L. Phylogenetic relationships of Cryptosporidium parasites based on the 70-Kilodalton Heat Shock Protein (HSP70) gene. Appl Eviron Microbiol. 2000; 66: 2385-2391.

Glaberman S, Sulaiman IM, Bern C, et al. A multilocus genotypic analysis of Cryptosporidium meleagridis. J Eukaryot Microbiol. 2001; Suppl: 19s-22s.

How to Cite
Ghaffari S, Kalantari N. A multi-locus study of cryptosporidium parasites isolated from patients living in iran, Malawi, Nigeria, the United kingdom, and Vietnam. IJPA. 9(1):79-.