Validation of droplet digital Polymerase Chain Reaction for the detection and absolute quantification of Taenia solium eggs in spiked soil samples

Maganira, Justine Daudi; Mwang'onde, Beda J; Kidima, Winifrida; Chacha, Mwita; Nkwengulila, Gamba; Höglund, Johan

doi:10.1016/j.actatropica.2019.105175

Cited by 8 publications

(18 citation statements)

References 28 publications

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“…Tyler, Ohio, USA). Then, about 10 g of the filtered soil from each household was transferred into a 50 ml Falcon tube for T. solium egg recovery using 15 ml of saturated zinc chloride solution (density 1.45 g/ml, Acros Organics, Geel, Belgium), basically in accordance with a protocol originally developed for detection of the related cestode Echinococcus multilocularis eggs in fox faeces [ 22 , 23 ], with minor modifications as described in 17 .…”

Section: Methodsmentioning

confidence: 99%

“…Initially, all 192 samples were screened using universal primers suggested by 3 , which amplify the mitochondrial cytochrome c oxidase subunit I ( cox1 ] gene from a range of cestodes and trematodes, in ddPCR EvaGreen Supermix (Bio-Rad Laboratories) [universal ddPCR assay) as described in 17 . The presence of T. solium DNA was then confirmed in all universal ddPCR assay positive reactions using a specific primers probe set in ddPCR Supermix for Probes with no dUTP (Bio-Rad Laboratories] (specific ddPCR probe assay) targeting the mitochondrial cox1 gene of T. solium .…”

Section: Methodsmentioning

confidence: 99%

“…The optimal annealing temperature for the universal ddPCR assay was set to 54°C [17]. For the ddPCR probe assay, the annealing temperature was determined to be 56°C by a thermal gradient ranging from 50°C to 60°C with 2 µl of template T. solium cysticerci DNA extracted from pigs [5].…”

Section: Dna Examinationmentioning

confidence: 99%

“…eggs, but these suffer from limitations [16]. It was not until recently, a droplet digital Polymerase Chain Reaction (ddPCR) technique was validated for the detection of parasite egg DNA from soil [17]. However, application of this technique when performed in soils collected from the field environment has not been done.…”

Section: Introductionmentioning

confidence: 99%

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Soil contamination by Taenia solium egg DNA in rural villages in Kongwa district, Tanzania

Maganira

Kidima

Chacha

et al. 2020

Infection Ecology & Epidemiology

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The presence ofTaenia solium DNA from eggs in soils around the households in four Tanzanian villages in Kongwa district were analysed in relation to seasonal fluctuations and infection risk implications. A total of 192 pooled soil samples from five sampling points per household were examined by droplet digital Polymerase Chain Reaction (ddPCR) from 96 pigkeeping households both during the dry and rainy seasons. The pooled samples were first processed by a flotation-double sieving technique, followed by screening for worm DNA employing universal primers targeting the mitochondrial cytochrome c oxidase subunit I (cox1) gene of human taeniid species and some other helminths. All DNA positive samples were later confirmed by a specific ddPCR probe assay targeting the mitochondrial cox1 gene of T. solium. A total of 17.2% (n = 33) samples were positive with the universal ddPCR, whereas T. solium DNA was confirmed by the specific ddPCR only in 3.1% (n = 3) of the surveyed households. The detection of T. solium DNA in this study spells out a low risk of exposure to T. solium eggs from contaminated household soil. Based on our results, ddPCR seems to be a promising technology for screening T. solium eggs in soil.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Dna Examinationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Soil contamination by Taenia solium egg DNA in rural villages in Kongwa district, Tanzania

Maganira

Kidima

Chacha

et al. 2020

Infection Ecology & Epidemiology

Self Cite

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show abstract

“…Over the past 2 decades, Tanzania's pig population, pork production, and pork consumption have all increased significantly (Kimbi et al 2015 ). For example, in rural areas in the southern highland regions, such as Mbeya, Rukwa, Songwe, Iringa, Ruvuma, and Njombe, at least one in every four households keeps pigs (Shonyela et al 2017 ; Kabululu et al 2018 ; Maganira et al 2018 , 2019a , b ; Openshaw et al 2018 ). These areas generate the majority of Tanzania's food crops (Kabululu et al 2018 ), and raising pigs is advantageous because most people feed their pigs with farm leftovers.…”

Section: Introductionmentioning

confidence: 99%

Socio-economic and health costs of porcine/human cysticercosis, neurocysticercosis and epilepsy to small-scale pig producers in Tanzania

Kayuni¹

2021

Bull Natl Res Cent

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Background Porcine cysticercosis, human cysticercosis, neurocysticercosis, and epilepsy pose a serious public health risk and lead to economic losses to the small-scale pig farmers in Tanzania’s rural areas. It also resulted in lowering the pig’s prices, high treatment expenses, lower labour productivity, and social discrimination. In Tanzania, small-scale pig producers are unaware of the methods that are used to manage and prevent the diseases associated with Taenia solium. Main body The main goal of the reviewed article was to investigate the socio-economic and health costs of porcine/human cysticercosis, neurocysticercosis, and epilepsy on small-scale pig producers in Tanzania. Several search engines yielded 80 peer-reviewed publications, 44 of which were considered to be relevant to the study. Taenia solium cysticercosis has been reported to cause huge financial losses in the management of pigs posing a burden in terms of vaccination and treatment. Conclusion The information on porcine/human cysticercosis, neurocysticercosis, and epilepsy is still insufficient in Tanzania. Poor habits, negative perceptions, and attitudes are the result of this misinformation. This review suggests that health education should be emphasized as one of the intervention methods for eliminating all forms of cysticercosis and epilepsy in Tanzania to promote and increase awareness about all forms of cysticercosis and epilepsy in rural areas.

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Diagnostic tools for the detection of taeniid eggs in different environmental matrices: A systematic review.

Saelens

Robertson

Gabriël

2022

Food and Waterborne Parasitology

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Validation of droplet digital Polymerase Chain Reaction for the detection and absolute quantification of Taenia solium eggs in spiked soil samples

Cited by 8 publications

References 28 publications

Soil contamination by Taenia solium egg DNA in rural villages in Kongwa district, Tanzania

Soil contamination by Taenia solium egg DNA in rural villages in Kongwa district, Tanzania

Socio-economic and health costs of porcine/human cysticercosis, neurocysticercosis and epilepsy to small-scale pig producers in Tanzania

Diagnostic tools for the detection of taeniid eggs in different environmental matrices: A systematic review.

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