Approaches to genotyping individual miracidia of Schistosoma japonicum

Xiao, Ning, Remais, Justin V., Brindley, Paul J., Qiu, Dong-Chuan, Carlton, Elizabeth J., Li, Rong-Zhi, Lei, Yang, and Blair, David (2013) Approaches to genotyping individual miracidia of Schistosoma japonicum. Parasitology Research, 112 (12). pp. 3991-3999.

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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.

Item ID: 30639
Item Type: Article (Research - C1)
ISSN: 1432-1955
Funders: NIH/NSF Ecology of Infectious Disease Program, National Institute for Allergy and Infectious Disease, Emory Global Health Institute
Projects and Grants: NIH/NSF Ecology of Infectious Disease Program (grant no. 0622743), National Institute for Allergy and Infectious Disease (grant no. K01AI091864 and R01AI068854)
Date Deposited: 18 Dec 2013 09:45
FoR Codes: 06 BIOLOGICAL SCIENCES > 0604 Genetics > 060411 Population, Ecological and Evolutionary Genetics @ 50%
11 MEDICAL AND HEALTH SCIENCES > 1108 Medical Microbiology > 110803 Medical Parasitology @ 50%
SEO Codes: 92 HEALTH > 9204 Public Health (excl. Specific Population Health) > 920404 Disease Distribution and Transmission (incl. Surveillance and Response) @ 100%
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