Evidence map›Paper›PMID 40345597›Full record

ArticleJournal of virological methods2025

Technical validation of a multiplex real-time PCR for combined detection of Rift Valley fever, chikungunya, Zika and dengue viruses.

Anne Hauner, Stijn Rogé, Veerle Vanlerberghe, Luciana Lepore, Fabrice Ndayisenga, Anselme Shyaka, Marjan Van Esbroeck, Silvia Situma, Carolyne Nasimiyu, Steve Ahuka-Mundeke and 5 more

Abstract readValidation Study
In one paragraph

Article in Journal of virological methods, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Evaluation of the NovaplexInternational journal of molecular sciences · 2026
    Article
  2. Integrating metagenomics and metatranscriptomics intoThe Journal of general virology · 2026
    Review
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

15 authors.

Anne HaunerDepartment of Biomedical Sciences, Unit of Virology, Institute of Tropical Medicine, Antwerp, Belgium. Electronic address: ahauner@itg.be.
Stijn RogéDRC office, Institute of Tropical Medicine, Antwerp, Belgium.
Veerle VanlerbergheDepartment of Public Health, Unit of Emerging Infectious Diseases, Institute of Tropical Medicine, Antwerp, Belgium.
Luciana LeporeDepartment of Public Health, Unit of Emerging Infectious Diseases, Institute of Tropical Medicine, Antwerp, Belgium.
Fabrice NdayisengaRwanda Agriculture and Animal Resources Development Board, Kigali, Rwanda.
Anselme ShyakaCenter for One Health, University of Global Health Equity, Butaro, Rwanda.
Marjan Van EsbroeckDepartment of Clinical Science, Institute of Tropical Medicine, Antwerp, Belgium.
Silvia SitumaWashington State University Global Health-Kenya, Nairobi, Kenya.
Carolyne NasimiyuWashington State University Global Health-Kenya, Nairobi, Kenya.
Steve Ahuka-MundekeNational Institute for Biomedical Research INRB, Kinshasa, Congo.
M Kariuki NjengaWashington State University, Pullman, USA.
Robert F BreimanEmory University, Atlanta, USA.
Justin MasumuNational Institute for Biomedical Research INRB, Kinshasa, Congo.
Daniel Mukadi-BamulekaRodolphe Mérieux INRB-Goma Laboratory, Goma, North Kivu, Congo; Service of Microbiology, Department of Medical Biology, Faculty of Medicine, University of Kinshasa, Kinshasa, Congo; Department of Virology, Institut National de Recherche Biomédicale, INRB, Kinshasa, Congo.
Kevin K AriënDepartment of Biomedical Sciences, Unit of Virology, Institute of Tropical Medicine, Antwerp, Belgium; Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium. Electronic address: karien@itg.be.

Funding

Emerging Infectious Diseases Research Center - East and Central AfricaU01AI151799 · NIAID · WASHINGTON STATE UNIVERSITY · PI M KARIUKI NJENGA · 2020 to 2026
$10.4M
Zoonotic and Emerging Infectious Diseases Training ProgramD43TW011519 · FIC · WASHINGTON STATE UNIVERSITY · PI Walter Godfrey Jaoko, Thumbi Mwangi · 2020 to 2026
$1.2M
FIC NIH HHS D43 TW011519NIAID NIH HHS U01 AI151799
6 · The paper itself

Abstract

Several arthropod-borne (arbo)-viruses have overlapping symptoms, insect vectors and geographical occurrence. With little known about the importance of arboviruses as cause of acute undifferentiated fever (AUF) in East and Central Africa (ECA), there is a clear need for a multiplex-PCR allowing for multi-pathogen surveillance. A multiplex real-time RT-PCR (RDCZ-multiplex) was developed and validated for the simultaneous detection of Rift Valley fever virus (RVFV), dengue virus 1-4 (DENV), chikungunya virus (CHIKV) and Zika virus (ZIKV). Phocine distemper virus (PDV) was added to the PCR as sample extraction control. Validation was conducted following the MIQE-guidelines using a panel of retrospective clinical samples and Quality Control for Molecular Diagnostics (QCMD, https://www.qcmd.org/en/) samples with the simplex-PCR as reference. These included samples from RVFV in animals (n = 19), DENV (n = 15), CHIKV (n = 11), ZIKV (n = 2) and YFV (n = 1, QCMD), and 14 negative endemic controls. Extractions and PCRs were done with commercially available kits. Some loss of sensitivity was observed at low target concentrations for RVFV, DENV1 and DENV4, when comparing the standard curves of simplex-PCRs with the multiplex-PCR. The limit of detection of the multiplex-PCR was 2064 copies/ml for CHIKV, 3587 copies/ml for DENV1, 30,249 copies/ml for ZIKV and 73 PFU/ml for RVFV. Specificity of the multiplex-PCRs was 100 %. For 12 out of 48 positive samples with high Cq values, RVFV (n = 7), CHIKV (n = 2), DENV1 (n = 2), YFV (n = 1), the multiplex-PCRs were negative. Although PCR sensitivity of the RDCZ-multiplex is slightly lower with low target concentrations, it offers a useful tool for molecular surveillance and clinical diagnosis for arboviruses for the ECA-region.

Indexed as

Arbovirus InfectionsDengue VirusMultiplex Polymerase Chain ReactionReal-Time Polymerase Chain ReactionAnimalsChikungunya FeverChikungunya virusDengueHumansRift Valley FeverRift Valley fever virusSensitivity and SpecificityZika VirusZika Virus InfectionArbovirusChikungunya virusDengue virusReal-time multiplex RT-PCRRift valley fever virusValidationZika virus

Identifiers

PMID40345597
PMCPMC12328903

What OpenQuestion holds

Textmetadata
LicenceCC BY-NC-ND
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Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.