Abstract
Arbovirus diagnostics on blood from donors and travelers returning from endemic areas is increasingly important for better patient management and epidemiological surveillance. We developed a flexible approach based on a magnetic field-enhanced agglutination (MFEA) readout to detect either genomes or host-derived antibodies. Dengue viruses (DENVs) were selected as models. For genome detection, a pan-flavivirus amplification was performed before capture of biotinylated amplicons between magnetic nanoparticles (MNPs) grafted with DENV probes and anti-biotin antibodies. Magnetization cycles accelerated this chaining process to within 5 min while simple turbidimetry measured the signal. This molecular MFEA readout was evaluated on 43 DENV RNA(+) and 32 DENV RNA(−) samples previously screened by real-time RT-PCR. The sensitivity and the specificity were 88.37% (95% CI, 78.76%–97.95%) and 96.87% (95% CI, 90.84%–100%), respectively. For anti-DENV antibody detection, 103 plasma samples from donors were first screened using ELISA assays. An immunological MFEA readout was then performed by adding MNPs grafted with viral antigens to the samples. Anti-DENV antibodies were detected with a sensitivity and specificity of 90.62% (95% CI, 83.50%–97.76%) and 97.44% (95% CI, 92.48%–100%), respectively. This adaptable approach offers flexibility to platforms dedicated to the screening of emerging infections.
Highlights
Dengue viruses (DENVs) belong to the genus Flavivirus that includes Zika, yellow fever, and West Nile viruses, and are transmitted to humans bitten by infected Aedes aegypti and Aedes albopictus mosquitoes [2,6]
We previously developed a multiplex approach based on a pan-flavivirus reverse transcription (RT)-PCR amplification combined with specific capture of amplified genomes on tetrathiolated probes grafted on microplates [11]
RNA levels were measured before shipment by the CNR using a reverse transcription (RT)-quantitative real-time PCR method (RT-qPCR)
Summary
The emergence and re-emergence of arthropod-borne viruses (arboviruses) belonging to the Flaviviridae, Togaviridae, Reoviridae, or Bunyaviridae families constitute a threat to global health following their spread throughout all continents [1,2,3]. Dengue, selected as a model in this study, is the most prevalent arbovirus disease affecting humans with an estimated. 390 million infections per year, 96 million of which are symptomatic, comprising 2 million severe with 21,000 fatal cases [4,5]. Dengue viruses (DENVs) belong to the genus Flavivirus that includes Zika, yellow fever, and West Nile viruses, and are transmitted to humans bitten by infected Aedes aegypti and Aedes albopictus mosquitoes [2,6]. Dengue disease is Microorganisms 2021, 9, 674.
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