Please use this identifier to cite or link to this item:
Authors: Koliopoulos, Philip
Kayange, Neema Mathias
Daniel, Tim
Huth, Florian
Gröndahl, Britta
Medina-Montaño, Grey Carolina
Pretsch, Leah
Klüber, Julia
Schmidt, Christian
Züchner, Antke
Ulbert, Sebastian
Mshana, Steven E.
Addo, Marylyn
Gehring, Stephan
Title: Multiplex-RT-PCR-ELISA panel for detecting mosquito-borne pathogens : Plasmodium sp. preserved and eluted from dried blood spots on sample cards
Online publication date: 24-Sep-2021
Language: english
Abstract: BACKGROUND Children are the most vulnerable group affected by malaria and other tropical, vector-borne diseases in low-resource countries. Infants presenting with acute onset fever represent a major sector of outpatient care in the Lake Victoria region. Misclassification and overuse of antibiotics and anti-malarial medications are consistent problems. Identifying the prevalent mosquito-borne pathogens in the region will reduce the prescription of non-indicated medicines. METHODS The literature was reviewed focusing on the mosquito-borne pathogens most prevalent in sub-Saharan Africa. Accordingly, an assay comprised of a multiplex-reverse transcriptase-polymerase chain reaction and an enzyme-linked immunosorbent assay (multiplex-RT-PCR-ELISA) was designed and validated in its ability to identify and differentiate nine human mosquito-borne pathogens including eight arboviruses and Plasmodium sp., the aetiologic agents of malaria. Blood samples obtained from 132 children suspected of having malaria were spotted and preserved on Whatman® 903 protein sample cards. Multiplex-RT-PCR-ELISA analysis was assessed and compared to results obtained by blood smear microscopy and the malaria rapid diagnostic test (RDT). RESULTS Nine out of nine pathogens were amplified specifically by the multiplex-RT-PCR-ELISA panel. Twenty-seven out of 132 paediatric patients presenting with acute fever were infected with Plasmodium sp., confirmed by multiplex-RT-PCR. The results of blood smear microscopy were only 40% sensitive and 92.8% specific. The malaria RDT, on the other hand, detected acute Plasmodium infections with 96.3% sensitivity and 98.1% specificity. The preservation of Plasmodium sp. in clinical sera and whole blood samples spotted on sample cards was evaluated. The duration of successful, sample card storage was 186 to 312 days. CONCLUSIONS Reliable, easy-to-use point of care diagnostic tests are a powerful alternative to laboratory-dependent gold standard tests. The multiplex-RT-PCR-ELISA amplified and identified nine vector-borne pathogens including Plasmodium sp. with great accuracy. Translation of improved diagnostic approaches, i.e., multiplex-RT-PCR-ELISA, into effective treatment options promises to reduce childhood mortality and non-indicated prescriptions.
DDC: 610 Medizin
610 Medical sciences
Institution: Johannes Gutenberg-Universität Mainz
Department: FB 04 Medizin
Place: Mainz
Version: Published version
Publication type: Zeitschriftenaufsatz
License: CC BY
Information on rights of use:
Journal: Malaria journal
Pages or article number: 66
Publisher: BioMed Central
Publisher place: London
Issue date: 2021
ISSN: 1475-2875
Publisher URL:
Publisher DOI: 10.1186/s12936-021-03595-4
Appears in collections:JGU-Publikationen

Files in This Item:
  File Description SizeFormat
koliopoulos_philip-multiplex-rt-p-20210922195641275.pdf1.97 MBAdobe PDFView/Open