Distinguishing Echinococcus granulosus sensu stricto genotypes G1 and G3 with confidence: A practical guide

Kinkar, Liina; Laurimäe, Teivi; Acosta‐Jamett, Gerardo; Andresiuk, Vanessa; Balkaya, İbrahim; Casulli, Adriano; Gasser, Robin B.; González, Luís Miguel; Haag, Karen Luisa; Zait, H.; Irshadullah, Malik; Jabbar, Abdul; Dj, Jenkins; Manfredi, Maria Teresa; Mirhendi, Hossein; M’rad, Selim; Rostami-Nejad, Mohammad; Oudni-M’rad, Myriam; Pierangeli, Nora Beatriz; Ponce-Gordo, Francisco; Rehbein, Steffen; Sharbatkhori, Mitra; Kia, Eshrat Beigom; Şimşek, Sami; Soriano, Silvia Viviana; Sprong, Hein; Šnábel, Viliam; Umhang, Gérald; Varcasia, Antonio; Saarma, Urmas

doi:10.1016/j.meegid.2018.06.026

Cited by 68 publications

(89 citation statements)

References 47 publications

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“…Only one case of G3 strain and 3 cases of G1/G3 strains were found in this study. In general, E. granulosus sensu stricto (G1-G3) are the predominant strains in CE cases throughout the world (7,8). Findings of the current study are in accordance with other reports from Iran, Turkey, and also the Middle East countries.…”

Section: Discussionsupporting

confidence: 90%

“…Apart from the G4 genotype, all other strains of E. granulosus have been identified as the cause of human CE. The genotypes G1 and G3 are the most common genotypes identified in livestock and human all over the world (7,8). The infection has been reported from all of the Middle Eastern countries and in the meantime, Iran and Turkey are considered as hyper-endemic areas for human CE (9)(10)(11).…”

Section: Introductionmentioning

confidence: 99%

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Comparative Genotyping of <i>Echinococcus granulosus</i> Infecting Livestock in Turkey and Iran

Barazesh¹,

Sarkari²,

Sarısu³

et al. 2019

Turkiye Parazitol Derg

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Amaç: Echinococcus granulosus, yaşam döngüsü paterni ve konak tiplerine göre çeşitlilik gösteren farklı suşlara sahiptir. Şimdiye kadar bu parazitin 10 genotipi, moleküler yöntemler kullanılarak tespit edilmiştir. Bu çalışmada, Türkiye ve İran'daki hayvanlarda E. granulosus metasestodlarının genotipik çeşitliliğinin değerlendirilmesi ve karşılaştırılması amaçlanmıştır. Yöntemler: İran'ın Doğu Azerbaycan eyaletindeki Bonab şehrindeki (30 koyun ve 30 sığır dahil olmak üzere 60 örnek) ve Türkiye'nin Van şehrindeki (15 koyun ve 15 sığır dahil olmak üzere 30 örnek) endüstriyel kesimhanelerinden hidatik kist ile enfekte Objective: Echinococcus granulosus contains a complex of different strains that represent diversity in the pattern of the life cycle and also their host types. So far 10 genotypes of this parasite have been identified, using molecular methods. The current study aimed to evaluate and compare the genotypic diversity of E. granulosus metacestodes from livestock of Turkey and Iran. Methods: A total of 90 livestock liver and lung organs infected with hydatid cyst from industrial slaughterhouses of Bonab Province in the East Azerbaijan Province in Iran (60 samples, including 30 sheep and 30 cattle) and Van Province in Turkey (30 samples, including 15 sheep and 15 cattle) were collected. DNA was extracted from the protoscolices or germinal layers and polymerase chain reaction (PCR) were utilized, targeting the partial mitochondrial cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase 1 (nad1) genes. PCR products were isolated from the electrophoresis gels and sequenced. The sequences were compared with each other, as well as with those related available sequences in the GenBank, using the BioEdit software and the BLAST algorithm. Finally, the phylogenetic trees were constructed by comparing sequences of cox1 and nad1 fragments, using the MEGA7 software and the maximum likelihood method. Results: All samples sequenced from Iran corresponded to the genotype G1 (100%). Among the samples from Turkey, 15 samples (78.9%) were identified as G1 while only one sample (5.3%) corresponded to the genotype G3 and 3 isolates (15.8%) were defined as genotypes G1/G3. Five distinct haplotypes were determined within the examined isolates from sheep and cattle in both countries and all isolates clustered in one group. Phylogenetic analysis revealed that the intra-species genetic variations were 0.0-0.6% and 0.0-1.4% for cox1 and nad1, respectively. Conclusion: The dominant genotype of E. granulosus sensu stricto of livestock in both countries was the G1 (sheep strain) genotype. Our findings indicate that the sheepdog cycle is the leading cycle of E. granulosus in these two areas. Hence, adopting regional common policies and bilateral cooperation helps to control the disease in livestock as well as in human in these two regions. Further study is required to compare the genetic diversity of human isolates of E. granulosus in these two countries.

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Section: Discussionsupporting

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Comparative Genotyping of <i>Echinococcus granulosus</i> Infecting Livestock in Turkey and Iran

Barazesh¹,

Sarkari²,

Sarısu³

et al. 2019

Turkiye Parazitol Derg

View full text Add to dashboard Cite

show abstract

“…Although the amplification of the nuclear CAL gene consents to identify E. granulosus s.s. G1-G3 genotype, by this, and other genes, it is not possible to differentiate between these two genotypes [44]. At the moment, there is no suitable method for the identification and differentiation among these genotypes G1 and G3, apart from sequencing of almost the entire mitochondrial genome or amplifying same mitochondrial markers genes [44]. Indeed, according to a recent study on the phylogenetic network the Nad5 gene fragment (680 bp) permitted to clear distinguish between the genotypes G1 and G3 instead, other mitochondrial marker genes, such as COX1 (366 bp) and/or NAD1 (471 bp) [12,13], are able to distinguish between G1 and G3 genotypes when associated to Nad5 [44].…”

Section: Discussionmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 98%

“…At the moment, there is no suitable method for the identification and differentiation among these genotypes G1 and G3, apart from sequencing of almost the entire mitochondrial genome or amplifying same mitochondrial markers genes [44]. Indeed, according to a recent study on the phylogenetic network the Nad5 gene fragment (680 bp) permitted to clear distinguish between the genotypes G1 and G3 instead, other mitochondrial marker genes, such as COX1 (366 bp) and/or NAD1 (471 bp) [12,13], are able to distinguish between G1 and G3 genotypes when associated to Nad5 [44]. To develop and validate a test is a procedure that lead to improve the performance of the methods and consists on evaluation of the feasibility, development and standardization, determination of the characteristics and a continuous monitoring maintenance and improvement of the assay [40].…”

Section: Discussionmentioning

confidence: 99%

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Validation of a one-step PCR assay for the molecular identification of Echinococcus granulosus sensu stricto G1–G3 genotype

et al. 2019

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The Italian National Reference Center for Echinococcosis (CeNRE, Sassari, Italy) set up a diagnostic protocol of "one-step-PCR" useful for the detection of E. granulosus sensu stricto (E.g.s.s.) and the identification of its genotype (G1-G3). The purpose of this work was to perform the validation of the "PCR E.g.s.s." method. The procedures were performed employing the criteria of the World Organization for Animal Health as well as of the Italian Accreditation Body (ACCREDIA) based on the Regulation UNI CEI EN ISO/IEC 17025. Positive DNA samples belonging to E. granulosus, E. ortleppi, E. multilocularis, E. canadensis species were used for the experiments. Analytical specificity evidenced primer pairs Cal (Calreticulin l gene of 1001 bp) with an specificity higher respect to Ef1 (Elongation-Factor 1 Alpha gene of 706 bp) and NAD (Dehydrogenase-subunit 1 gene of 219 bp). The analytical sensitivity presented the capability to detect a very low amount of parasite DNA corresponding to a concentration of 12.5 pg/µl; accuracy and precision related to the operator performance, along with repeatability and reproducibility, evidenced high concordance among results and demonstrated an excellent κ values of Cohen. According to the good performance related to the evaluated parameters, the method "PCR E.g.s.s." was suitable for the validation procedure, and consequently, to be undergone to the accreditation process. In conclusion, the results demonstrated an elevated robustness and reliable features of the "PCR E.g.s.s." able to perform a rapid diagnosis of E. granulosus in only "one step", hence, it is likely to avoid the sequencing step.

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First detection of zoonotic tapeworm Echinococcus granulosus sensu lato genotype G7 in continental Italy

et al. 2019

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Distinguishing Echinococcus granulosus sensu stricto genotypes G1 and G3 with confidence: A practical guide

Cited by 68 publications

References 47 publications

Comparative Genotyping of <i>Echinococcus granulosus</i> Infecting Livestock in Turkey and Iran

Comparative Genotyping of <i>Echinococcus granulosus</i> Infecting Livestock in Turkey and Iran

Validation of a one-step PCR assay for the molecular identification of Echinococcus granulosus sensu stricto G1–G3 genotype

First detection of zoonotic tapeworm Echinococcus granulosus sensu lato genotype G7 in continental Italy

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