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Approaches to genotyping individual miracidia of Schistosoma japonicum

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Abstract

Molecular genetic tools are needed to address questions as to the source and dynamics of transmission of the human blood fluke Schistosoma japonicum in regions where human infections have reemerged, and to characterize infrapopulations in individual hosts. The life stage that interests us as a target for collecting genotypic data is the miracidium, a very small larval stage that consequently yields very little DNA for analysis. Here, we report the successful development of a multiplex format permitting genotyping of 17 microsatellite loci in four sequential multiplex reactions using a single miracidium held on a Whatman Classic FTA indicating card. This approach was successful after short storage periods, but after long storage (>4 years), considerable difficulty was encountered in multiplex genotyping, necessitating the use of whole genome amplification (WGA) methods. WGA applied to cards stored for long periods of time resulted in sufficient DNA for accurate and repeatable genotyping. Trials and tests of these methods, as well as application to some field-collected samples, are reported, along with the discussion of the potential insights to be gained from such techniques. These include recognition of sibships among miracidia from a single host, and inference of the minimum number of worm pairs that might be present in a host.

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Acknowledgments

The authors wish to thank Kang Jun-xing, former General Director of the Sichuan Center for Disease Control and Prevention (Chengdu, People’s Republic of China) for his continued support and collaboration. The authors also wish to acknowledge Zhong Bo, Lu Ding, Ye Hong, Cui Lina, Chen Lin, Zhang Yi, and Meng Xian-hong of the Institute of Parasitic Diseases, Sichuan Center for Disease Control and Prevention for their support and assistance with the work described herein. We also thank county Anti-Schistosomiasis Control Station leaders and staff from the two counties included in this study for their hard work and hospitality during the fieldwork of sample collection conducted in 2007, 2008, and 2010. This work was supported in part by the NIH/NSF Ecology of Infectious Disease Program (grant no. 0622743), by the National Institute for Allergy and Infectious Disease (grant no. K01AI091864 and R01AI068854), and by the Emory Global Health Institute. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflict of interests

No author has any competing interests to declare.

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Author notes

  1. Ning Xiao

    Present address: National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Centre for Malaria, Schistosomiasis and Filariasis, Key Laboratory of Parasite and Vector Biology, Ministry of Health, Shanghai, 200025, People’s Republic of China

Authors and Affiliations

  1. Institute of Parasitic Diseases, Sichuan Center for Disease Control and Prevention, Chengdu, Sichuan, 610041, People’s Republic of China

    Ning Xiao, Dong-Chuan Qiu, Rong-Zhi Li & Yang Lei

  2. Department of Environmental Health, Rollins School of Public Health, Emory University, 1518 Clifton Rd. NE, Atlanta, GA, 30322, USA

    Justin V. Remais

  3. Department of Microbiology, Immunology and Tropical Medicine, George Washington University Medical Center, 2300 Eye Street, NW, Washington, DC, 20037, USA

    Paul J. Brindley

  4. Department of Environmental and Occupational Health, Colorado School of Public Health, University of Colorado, Anschutz Medical Campus, 13001 E. 17th Place, Aurora, CO, 80045, USA

    Elizabeth J. Carlton

  5. School of Marine and Tropical Biology, James Cook University, Townsville, QLD, 4811, Australia

    David Blair

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Correspondence to Ning Xiao.

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Xiao, N., Remais, J.V., Brindley, P.J. et al. Approaches to genotyping individual miracidia of Schistosoma japonicum . Parasitol Res 112, 3991–3999 (2013). https://doi.org/10.1007/s00436-013-3587-9

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  • DOI: https://doi.org/10.1007/s00436-013-3587-9

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