Isoenzyme profiles and phylogenetic analysis of Giardia duodenalis isolates from Iranian patients
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RESEARCH ARTICLE
Isoenzyme profiles and phylogenetic analysis of Giardia duodenalis isolates from Iranian patients Mohammad Rayani 1,2 & Ngah Zasmy Unyah 3 & Arghavan Vafafar 4 & Gholam Reza Hatam 5 Received: 21 January 2020 / Accepted: 7 July 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract The main objective of this study was to characterize the Giardia duodenalis isolates from Iranian patients in Fars Province, south of Iran by biochemical and molecular methods. Fifteen mass cultivated of G. duodenalis isolates in modified TYI-S-33 medium were analyzed using isoenzyme electrophoresis and PCR genotyping. Polyacrylamide gel electrophoresis (PAGE) of five different enzyme systems was used to characterize isolates: (i) glucose-6-phosphate dehydrogenase, (ii) glucose phosphate isomerase, (iii) malate dehydrogenase, (iv) malic enzyme, and (v) phosphoglucomutase. As well, a fragment of the SSUrDNA (292 bp) gene was amplified by PCR using the primers RH11 and RH4. The sequencing of the PCR products and phylogenetic tree were performed. The isoenzyme electrophoretic profiles divided fifteen G. duodenalis isolates into four zymodemes. G6PD, GPI, MDH, ME, and PGM enzyme systems showed 1, 2, 2, 3, and 3 enzyme pattern, respectively. G6PD isoenzyme pattern had the most homogeneity, while isoenzyme patterns of ME and PGM had the most heterogeneity in our study. Genotyping results indicated that the zymodemes 1–4 were categorized in assemblage A based on the SSU-rDNA gene. Phylogenetic analysis showed that all four zymodemes were distributed within the cluster of assemblage A. Our results indicated that both isoenzyme and DNA analyses were useful to characterize the isolates of Giardia and distinguishing various zymodemes and assemblages. It could be suggested that the genetic diversity among isoenzymes profiles of G. duodenalis may explain the variable clinical manifestations, pathogenicity, host response, drug susceptibility, and treatment efficacy of human giardiasis. Keywords Giardia duodenalis . Isoenzyme electrophoresis . Phylogenetic analysis . SSU-rDNA gene . Isoenzyme profiles
Introduction
Responsible Editor: Lotfi Aleya * Gholam Reza Hatam [email protected] 1
Persian Gulf Tropical Medicine Research Center, The Persian Gulf Biomedical Sciences Research Institute, Bushehr University of Medical Sciences, Bushehr, Iran
2
Department of Public Health, Faculty of Health and Nutrition, Bushehr University of Medical Sciences, Bushehr, Iran
3
Department of Medical Microbiology and Parasitology, Faculty of Medicine and Health Sciences, University Putra Malaysia, 43400 Selangor, Malaysia
4
Department of Parasitology and Mycology‚ School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
5
Basic Sciences in Infectious Diseases Research Center, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Genetic characterization between G. duodenalis strains has important clinical, diagnostic, and therapeutic implications of human giardiasis. It is not possible to
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