Blastocystis: Subtyping isolates using pyrosequencing™ technology

C.R. Stensvold; R.J. Traub; G. von Samson-Himmelstjerna; C. Jespersgaard; H.V. Nielsen; R.C.A. Thompson

doi:10.1016/j.exppara.2006.12.002

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Blastocystis: Subtyping isolates using pyrosequencing™ technology

Journal article

Peer reviewed

Blastocystis: Subtyping isolates using pyrosequencing™ technology

C.R. Stensvold, R.J. Traub, G. von Samson-Himmelstjerna, C. Jespersgaard, H.V. Nielsen and R.C.A. Thompson

Experimental Parasitology, Vol.116(2), pp.111-119

06/2007

DOI: https://doi.org/10.1016/j.exppara.2006.12.002

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Abstract

Blastocystis is a prevalent single-celled enteric parasite of unresolved clinical significance. Efforts based on molecular methodologies to establish whether pathogenicity is linked to specific isolates of the genetically diverse genus of Blastocystis have been scarce and so far yielded ambiguous results which can be difficult to interpret. To alleviate some of the problems related to unravelling the molecular epidemiology of Blastocystis infections we developed and evaluated a simple and high-throughput sequence analysis (SQA) pyrosequencing technique based on the detection of genotype-specific nucleotide polymorphisms in the 18S small subunit rRNA gene for a rapid and cost-effective post-PCR screening of Blastocystis genotypes. The method was effectively capable of genotyping 48/48 isolates positive by nested PCR in approximately one hour, and in 94% of the cases the isolate detected by PCR and pyrosequencing was also detected by one of two different PCR assays with subsequent dideoxy sequencing.

Details

Title: Blastocystis: Subtyping isolates using pyrosequencing™ technology
Authors/Creators: C.R. Stensvold (Author/Creator)
R.J. Traub (Author/Creator)
G. von Samson-Himmelstjerna (Author/Creator)
C. Jespersgaard (Author/Creator)
H.V. Nielsen (Author/Creator)
R.C.A. Thompson (Author/Creator)
Publication Details: Experimental Parasitology, Vol.116(2), pp.111-119
Publisher: Academic Press
Identifiers: 991005544517407891
Copyright: © 2006 Elsevier Inc. All rights reserved.
Murdoch Affiliation: School of Veterinary and Biomedical Sciences
Language: English
Resource Type: Journal article

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Collaboration types: Domestic collaboration; International collaboration
Citation topics: 1 Clinical & Life Sciences; 1.246 Diarrheal Diseases; 1.246.985 Cryptosporidium
Web Of Science research areas: Parasitology
ESI research areas: Microbiology