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210932

Algorithms based on PCR-RFLP of nad1 gene for genotyping of Echinococcus granulosus from human and animal isolates in Egypt

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Last updated: 22 Jan 2023

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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.

DOI

10.21608/puj.2021.94807.1131

Keywords

Algorithm, E. granulosus, Egypt, Genotyping, nad1, PCR-RFLP

Authors

First Name

Doaa

Last Name

Nassar

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Affiliation

Department of Medical Parasitology, Faculty of Medicine, Ain Shams University, Ramsis Street, Abbassia, 11591 Cairo, Egypt

Email

dr_doaa_ashraf@med.asu.edu.eg

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Orcid

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First Name

Ahmed

Last Name

Khalifa

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Affiliation

Department of Medical Parasitology, Faculty of Medicine, Ain Shams University, Ramsis Street, Abbassia, 11591 Cairo, Egypt

Email

ahmedkhalifaelsayed97@gmail.com

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Orcid

-

First Name

Hala

Last Name

Elwakil

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-

Affiliation

Department of Medical Parasitology, Faculty of Medicine, Ain Shams University, Ramsis Street, Abbassia, 11591 Cairo, Egypt

Email

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-

Orcid

-

First Name

Hayam

Last Name

Ezz Eldin

MiddleName

-

Affiliation

Department of Medical Parasitology, Faculty of Medicine, Ain Shams University, Ramsis Street, Abbassia, 11591 Cairo, Egypt

Email

hayamezz@hotmail.com

City

-

Orcid

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First Name

Hanan

Last Name

Abou-Seri

MiddleName

-

Affiliation

Department of Medical Parasitology, Faculty of Medicine, Ain Shams University, Ramsis Street, Abbassia, 11591 Cairo, Egypt

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Volume

14

Article Issue

3

Related Issue

29661

Issue Date

2021-12-01

Receive Date

2021-09-07

Publish Date

2021-12-01

Page Start

278

Page End

285

Print ISSN

1687-7942

Online ISSN

2090-2646

Link

https://puj.journals.ekb.eg/article_210932.html

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https://puj.journals.ekb.eg/service?article_code=210932

Order

6

Type

Original Article

Type Code

426

Publication Type

Journal

Publication Title

Parasitologists United Journal

Publication Link

https://puj.journals.ekb.eg/

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Article

Created At

22 Jan 2023