Nassar, D., Khalifa, A., Elwakil, H., Ezz Eldin, H., Abou-Seri, H. (2021). Algorithms based on PCR-RFLP of nad1 gene for genotyping of Echinococcus granulosus from human and animal isolates in Egypt. Parasitologists United Journal, 14(3), 278-285. doi: 10.21608/puj.2021.94807.1131
Doaa Nassar; Ahmed Khalifa; Hala Elwakil; Hayam Ezz Eldin; Hanan Abou-Seri. "Algorithms based on PCR-RFLP of nad1 gene for genotyping of Echinococcus granulosus from human and animal isolates in Egypt". Parasitologists United Journal, 14, 3, 2021, 278-285. doi: 10.21608/puj.2021.94807.1131
Nassar, D., Khalifa, A., Elwakil, H., Ezz Eldin, H., Abou-Seri, H. (2021). 'Algorithms based on PCR-RFLP of nad1 gene for genotyping of Echinococcus granulosus from human and animal isolates in Egypt', Parasitologists United Journal, 14(3), pp. 278-285. doi: 10.21608/puj.2021.94807.1131
Nassar, D., Khalifa, A., Elwakil, H., Ezz Eldin, H., Abou-Seri, H. Algorithms based on PCR-RFLP of nad1 gene for genotyping of Echinococcus granulosus from human and animal isolates in Egypt. Parasitologists United Journal, 2021; 14(3): 278-285. doi: 10.21608/puj.2021.94807.1131
Algorithms based on PCR-RFLP of nad1 gene for genotyping of Echinococcus granulosus from human and animal isolates in Egypt
Department of Medical Parasitology, Faculty of Medicine, Ain Shams University, Ramsis Street, Abbassia, 11591 Cairo, Egypt
Abstract
Background: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) is a simple rapid method for genotyping of Echinococcus garnulosus sensu lato (E. granulosus s. l.) in developing countries. Construction of algorithms based on PCR-RFLP using two restriction enzymes would be useful to study the genetic diversity of the parasite and would help in differentiation between ambiguous genotypes. Objective: The goal of the present work was to develop algorithms based on RFLP of nicotinamide adenine dinucleotide dehydrogenase (NADH) subunit 1 (nad1) sequences of reference genotypes of E. granulosus retrieved from GenBank for genotyping of human and animal isolates of E. granulosus in Egypt. Subjects and Methods: Retrieved nad1 sequences of reference genotypes were digested in silico individually with two restriction enzymes; Haemophilus influenza (HinfI) and Haemophilus aegyptius (HaeIII). The constructed PCR-RFLP algorithms were used for genotyping of 50 human and animal isolates (19 human, 23 camels and 8 pigs) analyzed by PCR-RFLP. To confirm the validity PCR-RFLP algorithms, samples corresponding to determined and undetermined genotypes as inferred from the algorithms were sequenced. Results: Utilizing PCR-RFLP and sequencing revealed that except for two cases (12.5%) which were typed as G1 among humans and one case as G5 in pigs (12.5%), G6 was the commonest genotype among human, camel and pig isolates collected. Conclusion: The algorithms based on PCR-RFLP of nad1 are valuable tools for genotyping of E. granulosuss. l. especially with HinfI RFLP algorithm. Sequencing is still needed to reveal the genotypes of undetermined or ambiguous isolates.
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