Detection of the Malaria causing Plasmodium Parasite in Saliva from Infected Patients using Topoisomerase I Activity as a Biomarker
| Autor: | Rella Manego Zoleko, Michael Ramharter, Anne Katrine Wrist Simonsen, Eskild Petersen, Rodrigo Labouriau, Lilian Endame, Anna Klicpera, Yi-Ping Ho, Johannes Mischlinger, Søren Fjelstrup, Ronja Neher, Felix Lötsch, Marianne Smedegaard Hede, Magnus Stougaard, Mirjam Groger, Ghyslain Mombo-Ngoma, Birgitta R. Knudsen, Luzia Veletzky |
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| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Saliva Plasmodium falciparum Protozoan Proteins lcsh:Medicine 02 engineering and technology Biology Article 03 medical and health sciences chemistry.chemical_compound Biotin Tandem repeat parasitic diseases medicine Humans Malaria Falciparum lcsh:Science Multidisciplinary Topoisomerase lcsh:R DNA 021001 nanoscience & nanotechnology medicine.disease Virology 030104 developmental biology Blue colored chemistry DNA Topoisomerases Type I Rolling circle replication biology.protein lcsh:Q Colorimetry 0210 nano-technology Malaria Biomarkers |
| Zdroj: | Hede, M S, Fjelstrup, S, Lötsch, F, Zoleko, R M, Klicpera, A, Groger, M, Mischlinger, J, Endame, L, Veletzky, L, Neher, R, Simonsen, A K W, Petersen, E, Mombo-Ngoma, G, Stougaard, M, Ho, Y-P, Labouriau, R, Ramharter, M & Knudsen, B R 2018, ' Detection of the Malaria causing Plasmodium Parasite in Saliva from Infected Patients using Topoisomerase I Activity as a Biomarker ', Scientific Reports, vol. 8, no. 1, pp. 4122 . https://doi.org/10.1038/s41598-018-22378-7 Scientific Reports Scientific Reports, Vol 8, Iss 1, Pp 1-12 (2018) |
| DOI: | 10.1038/s41598-018-22378-7 |
| Popis: | Malaria is among the major threats to global health with the main burden of disease being in rural areas of developing countries where accurate diagnosis based on non-invasive samples is in high demand. We here present a novel molecular assay for detection of malaria parasites based on technology that may be adapted for low-resource settings. Moreover, we demonstrate the exploitation of this assay for detection of malaria in saliva. The setup relies on pump-free microfluidics enabled extraction combined with a DNA sensor substrate that is converted to a single-stranded DNA circle specifically by topoisomerase I expressed by the malaria causing Plasmodium parasite. Subsequent rolling circle amplification of the generated DNA circle in the presence of biotin conjugated deoxynucleotides resulted in long tandem repeat products that was visualized colorimetrically upon binding of horse radish peroxidase (HRP) and addition of 3,3′,5,5′-Tetramethylbenzidine that was converted to a blue colored product by HRP. The assay was directly quantitative, specific for Plasmodium parasites, and allowed detection of Plasmodium infection in a single drop of saliva from 35 out of 35 infected individuals tested. The results could be determined directly by the naked eye and documented by quantifying the color intensity using a standard paper scanner. |
| Databáze: | OpenAIRE |
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