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.
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Data availability
Representative sequences obtained in this study were deposited in GenBank under the accession numbers OM780323–OM780326, OM791700–OM791701 and OM791622–OM791631.
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Acknowledgements
The authors thank all of the patients and field workers who participated in the study.
Funding
This study was funded by National Institute of Infectious Diseases (NIID), Tokyo, Japan.
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The authors confirm contribution to the article as follows: Sanjib K. Sardar: Conceptualization, methodology, data curation, writing original draft, revision of the manuscript. Seiki Kobayashi: Visualization, investigation, provide E. moshkovskii strain. Koushik Das: Formal analysis, validation. Yumiko Saito-Nakano: Validation, investigation. Shanta Dutta: Project administration. Tomoyashi Nozaki: Validation. Sandipan Ganguly: Conceptualization, formal analysis, funding acquisition, validation, visualization, review and editing, investigation, supervision.
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The protocol of this study was approved by the Institutional Human Ethics Committee of ICMR-National Institute of Cholera and Enteric Diseases, Kolkata. Informed consent for participation was obtained from every patient. In case of children the consent was obtained from their parents.
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The authors declare no competing interests.
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Sardar, S.K., Kobayashi, S., Das, K. et al. Development of a simple PCR–RFLP technique for detection and differentiation of E. histolytica, E. dispar and E. moshkovskii. Parasitol Res 122, 139–144 (2023). https://doi.org/10.1007/s00436-022-07706-1
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DOI: https://doi.org/10.1007/s00436-022-07706-1