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Use of PCR Amplification of tRNA Gene-Linked Short Tandem Repeats for Genotyping Entamoeba histolytica

Ibne Karim M. Ali, Mehreen Zaki , and C. Graham Clark^*

Department of Infectious and Tropical Diseases, London School of Hygiene and Tropical Medicine, Keppel Street, London WC1E 7HT, United Kingdom

Received 12 April 2005/ Returned for modification 25 August 2005/ Accepted 15 September 2005

We have developed a reliable method for PCR-based genotyping of Entamoeba histolytica based on variation in the numbers of short tandem repeats that are linked to tRNA genes in this species. Species-specific primer pairs were designed that differentiate E. histolytica from E. dispar as well as that reveal intraspecies PCR product length polymorphisms. The primers were tested with samples from different parts of the world, and DNA was extracted from cultured cells as well as liver abscess pus and feces by various methods. We now have the tools necessary to investigate a possible link between parasite genotype and the outcome of infection with Entamoeba histolytica, as well as other aspects of the organism's epidemiology.

Supplemental material for this article may be found at http://jcm.asm.org/.

Present address: Department of Microbiology, Stanford University School of Medicine, Stanford, CA 94305-5107.

Present address: School of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT, United Kingdom.

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