Restriction enzyme digestion analysis of PCR-amplified DNA of Blastocystis hominis isolates

Init, I.; Foead, A.L.; Fong, M.Y.; Yamazaki, H.; Rohela, M.; Yong, H.S.; Mak, J.W. (2007) Restriction enzyme digestion analysis of PCR-amplified DNA of Blastocystis hominis isolates. Southeast Asian Journal of Tropical Medicine and Public Health, 38 (6). pp. 991-7. ISSN 0125-1562

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    Genomic DNA of Blastocystis isolates released into 0.1 Triton X-100 was suitable for amplification and yielded similar results as the genomic DNA extracted with standard kit. The specific B. hominis primers (BH1: GCT TAT CTG GTT GAT CCT GCC AGT and BH2: TGA TCC TTC CGC AGG TTC ACC TAC A) successfully produced the PCR product of about 1,770 bp with all the 7 Blastocystis isolates tested. The restriction fragment length polymorphism (RFLP) patterns yielded by 13 out of 25 restriction endonucleases showed that the 7 isolates could be grouped into 4 subgroups: subgroup-1 consisted of isolate C; subgroup-2 of isolates H4 and H7; subgroup-3 of isolates KP1, Y51 and M12; and subgroup-4 of isolate 27805. The differences between subgroups manifested as clear-cut RFLP patterns. A common band of 230 bp was revealed by Eco R1 in all the Blastocystis isolates tested. The band of about 180 bp was revealed by Alu I, differentiated symptomatic from asymptomatic isolates of this parasite, and might indicate the pathogenicity of this parasite.

    Item Type: Article
    1. Init, I.(Department of Parasitology, Faculty of Medicine Building, University of Malaya, 50603 Kuala Lumpur, MALAYSIA)
    2. Foead, A.L.
    3. Fong, M.Y.
    4. Yamazaki, H.
    5. Rohela, M.
    6. Yong, H.S.
    7. Mak, J.W.
    Journal or Publication Title: Southeast Asian Journal of Tropical Medicine and Public Health
    Additional Information: Init, I Foead, A L Fong, M Y Yamazaki, H Rohela, M Yong, H S Mak, J W eng Research Support, Non-U.S. Gov't Thailand 2008/07/11 09:00 Southeast Asian J Trop Med Public Health. 2007 Nov;38(6):991-7.
    Uncontrolled Keywords: Animals Blastocystis Infections/parasitology Blastocystis hominis/*genetics/isolation & purification DNA, Protozoan/analysis Endonucleases/*genetics Humans Molecular Sequence Data *Polymerase Chain Reaction Polymorphism, Restriction Fragment Length/genetics Sequence Analysis, DNA
    Subjects: R Medicine
    Divisions: Faculty of Medicine
    Depositing User: Mr Jenal S
    Date Deposited: 10 Dec 2012 10:45
    Last Modified: 10 Dec 2012 10:45

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