Development of real-time PCR assay for simultaneous detection and genotyping of cystic echinococcosis in humans and livestock

Ahmed, Mohamed E.; Eldigail, Mawahib; Grobusch, Martin P.; Aradaib, Imadeldin E.

doi:10.12980/apjtd.7.2017d7-213

Cited by 3 publications

(4 citation statements)

References 33 publications

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“…Cystic echinococcosis (CE) is one of the most widespread parasitic zoonotic diseases in the world. Identification of the cyst genotype would be beneficial for prevention and control of the disease ( Ahmed et al, 2017 ). Four species of Echinococcus were recognized until a few years ago, namely, E. granulosus , E. multilocularis , E. vogeli , and E. oligarthus ( Thompson & McManus, 2002 ).…”

Section: Introductionmentioning

confidence: 99%

Genetic characterization of hydatid cysts of different intermediate hosts

et al. 2020

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SummaryCystic echinococcosis is an important cosmopolitan parasitic zoonosis that causes public health and economic problems in Egypt. The present study was undertaken to identify genotypes of hydatid cyst (HC) DNA isolated from different animal isolates and to identify the genotype of secondary hydatid cysts (HCs) developed in rabbits experimentally infected with camel HC for detection of any genetic mutation. In the present study, we extracted DNA from the germinal layers of 8 HCs collected from 3 camels, 1 cattle, 1 sheep and 3 donkeys in addition to 3 secondary HCs collected from rabbits experimentally infected with camel HC. PCR amplification of the ITS1 gene of all examined samples showed an amplified DNA band at 1115 bp. The partial nucleotide sequences of the ITS1 gene of all isolates were aligned and compared with the reference sequences of the genotypes G1–G8 in GenBank. The camel and rabbit samples were identified as Echinococcus canadensis genotype 6 (G6), while the cattle and sheep samples belonged to E. granulosus sensu stricto (G1). The donkey isolates belonged to E. equines (G4). Alignment of the ITS1 partial nucleotide sequences of the camel HCs and rabbit secondary HCs isolates with the G6 partial nucleotide sequence in GenBank was performed. Both camel HCs and rabbit secondary HCs isolates exhibited the same sequence identity matrix, which indicated the absence of mutation in the rabbit secondary HCs. It can be concluded that camel and rabbit samples were identified as E. canadensis (G6), the cattle and sheep samples belonged to E. granulosus sensu stricto (G1) and donkey isolates belonged to E. equines (G4). No mutation occurred during HCs transmission from camel to rabbit.

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Section: Introductionmentioning

confidence: 99%

Genetic characterization of hydatid cysts of different intermediate hosts

et al. 2020

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“…species were reported [ 34 ] while in Sudan, E . canadensis (G6/7) was the only species detected in 14 cases [ 41 – 43 ]. No other species (e.g.…”

Section: Resultsmentioning

confidence: 99%

“…A total of 465 human cysts were molecularly investigated, of which 443 (95.3%) were identified as E. PLOS NEGLECTED TROPICAL DISEASES [24,39,40]. In Tanzania, two E. granulosus s.s. species were reported [34] while in Sudan, E. canadensis (G6/7) was the only species detected in 14 cases [41][42][43]. No other species (e.g.…”

Section: Plos Neglected Tropical Diseasesmentioning

confidence: 99%

Epidemiology of Echinococcus granulosus sensu lato in the Greater Horn of Africa: A systematic review

Aregawi,

Levecke,

Ashenafi

et al. 2024

PLoS Negl Trop Dis

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Background Cystic echinococcosis (CE) is a neglected zoonotic disease that is caused by Echinococcus granulosus sensu lato (s.l.), the life cycle of which involves multiple hosts. We conducted a systematic review (SR) on E. granulosus s.l. in the Greater Horn of Africa (GHA), to provide a picture of its recent epidemiology across all hosts. Methods For this SR, conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement, five electronic databases, as well experts in the region were consulted to retrieve records published between 2000 and 2022, reporting the presence of E. granulosus s.l. infections in any natural host in the GHA (Djibouti, Eritrea, Ethiopia, Kenya, Sudan, Somalia, South Sudan, Tanzania and Uganda). Principal findings A total of 247 records were retained, describing the presence of E. granulosus s.l. throughout the GHA, except for Djibouti. Only few population surveys on human CE were conducted in the area, with the prevalence ranging between 0.3 and 11.3%. In animals, the reported prevalence ranged up to 61.6% in camels, 88.4% in cattle; 65.2% in goats, 9.9% in pigs, 67.8% in sheep and 94.5% in dogs. In addition, E. granulosus s.l. was also reported in wildlife. A total of five species were reported in the different hosts, namely E. granulosus sensu stricto (G1, G3, GOmo), E. canadensis (G6/7), E. ortleppi (G5), E. felidis, and E. equinus (G4). Conclusions We confirm that E. granulosus s.l. is prevalent throughout the GHA. Nevertheless, despite our efforts to screen grey literature, an accurate assessment of the epidemiology in GHA remains challenging, due to the lack of combined host, in-depth risk factor and behavioural studies, as well as the wide diversity in subpopulations studied and diagnostic tools used. Interdisciplinary and transboundary partnerships would be essential for the design of effective control strategies, tuned to the GHA setting.

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“…Several serological examination techniques have been developed for the detection of E. granulosus. However, these methods have low sensitivity and cross-reactivity is likely to outcome with other taeniid cestodes [6,8].…”

Section: Introductionmentioning

confidence: 99%

Development of Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick Assay for a Rapid and Sensitive Detection of Cystic Echinococcosis in Livestock in Kenya

Badoul¹,

Kagira

Ng’ong’a

et al. 2022

Journal of Tropical Medicine

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Background. Cystic echinococcosis is a zoonotic disease caused by the metacestode stage of Echinococcus granulosus and occurs worldwide, causing considerable economic losses and public health problems. The currently available methods for the diagnosis of animal hydatidosis are time-consuming and require well-equipped laboratories which make them incompatible with testing in resource-poor settings. This study developed and evaluated a rapid, more sensitive, and specific loop-mediated isothermal amplification combined with a lateral flow dipstick assay for the rapid and sensitive detection of cystic echinococcosis. Results. In this study, a specific primer set and FITC-labeled probe targeting the conserved region of the NADH-1 gene were designed. The LAMP reaction was performed at 60°C for 40 minutes, and the amplification products were successfully visualized by LFD strips. The analytical sensitivity of LAMP-LFD was determined using 10-fold serial dilutions of E. granulosus DNA. The minimal concentration detected was 10 fg/μl, and no cross-reactivity was observed with DNA extracted from Taenia solium, Taenia saginata, and Fasciola hepatica. The ability of the developed LAMP-LFD assay to detect cystic echinococcosis was further evaluated with 62 cyst samples from slaughtered cattle in Juja Abattoir, Kiambu County, Kenya. The LAMP-LFD was able to detect 59/62 (95.2%, 95% CI 0.87–0.98) as positive samples of E. granulosus compared to 53/62 (85.5%, 95% CI 0.75–0.92) by nested PCR assay. Conclusion. Our results indicated that the developed LAMP-LFD technique was more sensitive than the nested PCR assay, rapid, and easy to perform with a simple visual detection of products. Therefore, it could be an important point-of-care diagnostic tool for cystic echinococcosis.

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Development of real-time PCR assay for simultaneous detection and genotyping of cystic echinococcosis in humans and livestock

Cited by 3 publications

References 33 publications

Genetic characterization of hydatid cysts of different intermediate hosts

Genetic characterization of hydatid cysts of different intermediate hosts

Epidemiology of Echinococcus granulosus sensu lato in the Greater Horn of Africa: A systematic review

Development of Loop-Mediated Isothermal Amplification Combined with Lateral Flow Dipstick Assay for a Rapid and Sensitive Detection of Cystic Echinococcosis in Livestock in Kenya

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