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Updated: Aug 23, 2025
Development of a Negative Selectable Marker for Entamoeba histolytica
Published on: December 12, 2010
Development of a simple PCR-RFLP technique for detection and differentiation of E. histolytica, E. dispar and E.
Sanjib K Sardar1, Seiki Kobayashi2, Koushik Das1,3
1Division of Parasitology, ICMR-National Institute of Cholera and Enteric Diseases (ICMR-NICED), Kolkata, India.
Abstract:
Epidemiological studies on amoebic infections are complicated by morphological overlap between the pathogenic E. histolytica, the commensal E. dispar and the amphizoic E. moshkovskii, necessitating molecular identification. The present study developed a simple and economical 18S PCR-RFLP method for the simultaneous detection and differentiation of the three species. PCR products were differentiated by Tat1 restriction digestion generating three different RFLP patterns. Validation was conducted by screening 382 faecal samples from human patients from Kolkata, India, hospitalized for diarrhoea. Analysis indicated that the PCR-RFLP could successfully differentiate between the three species and was confirmed by sequence analysis. This method could prove useful for clinical and epidemiological studies of amoebiasis.
