Parasitology Research

, Volume 112, Issue 12, pp 3991–3999 | Cite as

Approaches to genotyping individual miracidia of Schistosoma japonicum

  • Ning Xiao
  • Justin V. Remais
  • Paul J. Brindley
  • Dong-Chuan Qiu
  • Elizabeth J. Carlton
  • Rong-Zhi Li
  • Yang Lei
  • David Blair
Original Paper

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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Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Ning Xiao
    • 1
    • 6
  • Justin V. Remais
    • 2
  • Paul J. Brindley
    • 3
  • Dong-Chuan Qiu
    • 1
  • Elizabeth J. Carlton
    • 4
  • Rong-Zhi Li
    • 1
  • Yang Lei
    • 1
  • David Blair
    • 5
  1. 1.Institute of Parasitic Diseases, Sichuan Center for Disease Control and PreventionChengduPeople’s Republic of China
  2. 2.Department of Environmental Health, Rollins School of Public HealthEmory UniversityAtlantaUSA
  3. 3.Department of Microbiology, Immunology and Tropical MedicineGeorge Washington University Medical CenterWashingtonUSA
  4. 4.Department of Environmental and Occupational Health, Colorado School of Public HealthUniversity of ColoradoAuroraUSA
  5. 5.School of Marine and Tropical BiologyJames Cook UniversityTownsvilleAustralia
  6. 6.National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, WHO Collaborating Centre for Malaria, Schistosomiasis and FilariasisKey Laboratory of Parasite and Vector Biology, Ministry of HealthShanghaiPeople’s Republic of China

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