Fig. 1 Schematic and representative spectra of Ebola, Lassa, and malaria detection using SERS nanotag technology. (A) Schematic of BD’s SERS nanotag technology. (B) Representative spectra from three SERS nanotags used in a sandwich immunoassay to detect Ebola, Lassa, and malaria histidine-rich protein 2 (HRP2) antigen. (C) Schematic of the HNW sandwich immunoassay using the SERS nanotags.
Fig. 2 Titration curves of singleplex assays and inclusivity testing in blood. Titration curve of (A) malaria (P. falciparum) HRP2 recombinant antigen, (B) live Ebola Kikwit virus, and (C) live Lassa Josiah virus. (D) Malaria inclusivity testing using the singleplex HRP2 assay with malaria-negative whole blood samples, P. vivax serum samples, and P. falciparum whole blood samples. (E) Ebola inclusivity testing using the singleplex assay with high concentrations of various strains and species of EBOV and (F) Lassa inclusivity testing using the singleplex assay with high concentrations of various LASVs. Figure results of assays run in triplicate; error bars reflect SD.
Fig. 3 Triplex assay tested with various antigens spiked. Signal seen from the EBOV, Lassa, and malaria assays is shown in blue, orange, and gray, respectively. Main figure shows signal generated after addition of ZIKV (MR766), ZIKV (PRVABC59), or DENV2 at 1 × 106 PFU/ml or recombinant Lassa NP, EBOV VP40, or P. falciparum HRP2 antigen to blood samples. Each bar represents an average of three experimental replicates; error bars represent SD.
Fig. 4 Assay testing on samples from EBOV-infected NHPs. (A) Time course of Ebola signals from blood from two different NHPs infected with Ebola Makona. Dashed line illustrates results from assays done on blood from one EBOV-infected NHP. Solid line illustrates results from assays done on blood from a second EBOV-infected NHP. Each point represents an average of three experimental replicates; error bars represent SD. (B) Triplex assay testing of NHP blood. Individual NHPs are represented on the x axis, one uninfected control (Neg) and nine late-stage Ebola Makona infected (1 to 9). Y axis shows assay signal from each of three triplex assays—EBOV HNW assay (blue diamonds), LASV HNW assay (orange triangle), and malaria HNW assay (gray circles).
Fig. 5 Results from clinical testing of the EBOV and malaria virus HNW assays. Boxplots of (A) assay signals from Ebola-positive (pos) and Ebola-negative (neg) samples. (B) Assay signals from malaria-positive and malaria-negative samples. (C and D) Corresponding ROC curves for Ebola and malaria, respectively. Area under the curve (AUC) for each ROC curve is shown in the figure, along with associated 95% confidence intervals (CIs).
- Table 1 Ebola and malaria status for the 586 clinical specimens at the Senegalese (276 samples) and Guinean (310 samples) field sites.
Site Known Ebola positive
(PCR)Known Ebola negative
(PCR)Known
malaria positive
(rapid assay and
microscopy)Known malaria
negative
(rapid assay and
microscopy)Total Guinea 100 90 52 68 310 Senegal 0 0 111 165 276 Total 100 90 163 233 586
Supplementary Materials
www.sciencetranslationalmedicine.org/cgi/content/full/10/471/eaat0944/DC1
Fig. S1. Schematic demonstrating the HNW assay workflow.
Fig. S2. Determination of assay sensitivity for Ebola, Lassa, and malaria HNW assays using recombinant antigens in assay buffer.
Fig. S3. Ebola inclusivity testing using the triplex assay with high concentrations of Sudan virus and Bundibugyo virus and a negative control.
Fig. S4. Signals obtained from recombinant antigens spiked into matched human blood, plasma, or serum that was provided by a single donor.
Fig. S5. Signal intensities for malaria and Ebola antigens spiked into duplex and triplex assay formats using blood as a test medium.
Fig. S6. ROC curves for the HNW malaria assay.
Fig. S7. Example SERS spectra from a clinical sample assay that indicated a single infectious agent.
Fig. S8. Example SERS spectra from a clinical sample assay that indicated a co-infection with malaria and EBOV.
Table S1. Reference and passage information for viruses used in the NHP model.
Table S2. Raw data for experiments where N < 20.
Additional Files
The PDF file includes:
- Fig. S1. Schematic demonstrating the HNW assay workflow.
- Fig. S2. Determination of assay sensitivity for Ebola, Lassa, and malaria HNW assays using recombinant antigens in assay buffer.
- Fig. S3. Ebola inclusivity testing using the triplex assay with high concentrations of Sudan virus and Bundibugyo virus and a negative control.
- Fig. S4. Signals obtained from recombinant antigens spiked into matched human blood, plasma, or serum that was provided by a single donor.
- Fig. S5. Signal intensities for malaria and Ebola antigens spiked into duplex and triplex assay formats using blood as a test medium.
- Fig. S6. ROC curves for the HNW malaria assay.
- Fig. S7. Example SERS spectra from a clinical sample assay that indicated a single infectious agent.
- Fig. S8. Example SERS spectra from a clinical sample assay that indicated a co-infection with malaria and EBOV.
- Table S1. Reference and passage information for viruses used in the NHP model.
Other Supplementary Material for this manuscript includes the following:
- Table S2 (Microsoft Excel format). Raw data for experiments where N < 20.
