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

Ithoi, Init and Foead, A.L. and Fong, Mun Yik and Yamazaki, H. and Rohela, M. and Yong, Hoi Sen and Mak, J.W. (2007) Restriction enzyme digestion analysis of PCR-amplified DNA of Blastocystis hominis isolates. The Southeast Asian Journal of Tropical Medicine and Public Health, 38 (6). 991 - 997. ISSN 0125-1562,

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Abstract

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 27B05. 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
Funders:	UNSPECIFIED
Additional Information:	Correspondence: Dr Init Ithoi, Department of Parasitology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. Fax: 603-79674754 E-mail: init@um.edu.my
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:	Fong Mun Yik
Date Deposited:	14 Jun 2012 01:27
Last Modified:	26 Aug 2020 07:09
URI:	http://eprints.um.edu.my/id/eprint/3264

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