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, (), -. doi: 10.21608/puj.2021.94807.1131

Doaa Ashraf Nassar; Ahmed Khalifa El Sayed Khalifa; Hala Salah ElWakil; Hayam Mohamed 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, , , 2021, -. 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, (), pp. -. 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; (): -. doi: 10.21608/puj.2021.94807.1131

Articles in Press, Accepted Manuscript, Available Online from 25 October 2021   Document Type: Original Article DOI: 10.21608/puj.2021.94807.1131Authors Doaa Ashraf Nassar1; Ahmed Khalifa El Sayed Khalifa2; Hala Salah ElWakil3; Hayam Mohamed Ezz Eldin4; Hanan Abou-Seri 51Parasitology department Faculty of Medicine Ain Shams university2Student, Faculty of Medicine, Ain Shams University3Parasitology Dept., Facutly of Medicine, Ain Shams Univ.4medical parasitology department faculty of medicine Ain shams university5Medical Parasitology, Faculty of Medicine, Ain Shams University, Cairo, EgyptAbstractBackground: 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. granulosus s. l. especially with HinfI RFLP algorithm. Sequencing is still needed to reveal the genotypes of undetermined or ambiguous isolates.Keywords algorithm; E. granulosus; Egypt; genotyping; nad1; PCR-RFLP Main Subjects Molecular Biology

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