Highly Sensitive and Selective Direct Detection of Zika Virus Particles in Human Bodily Fluids for Accurate Early Diagnosis of Infection

Abstract

Zika virus (ZIKV) is an arbovirus that caused widespread panic beginning in 2015 in northeastern Brazil due to the threatening link between infection and fetal abnormalities such as microcephaly, spontaneous abortions, and stillbirths. Since the epidemic began, the virus has been further investigated, unveiling that the long-term dangers of ZIKV infection go beyond fetal neurological impairment. Characterization of the active infection has proven difficult as only 20% of infected individuals are symptomatic. Additionally, ZIKV is often misdiagnosed due to serological cross-reactivity with similar flaviviruses such as dengue, yellow fever, and West Nile. To date, there is no approved vaccine or therapy against ZIKV, highlighting the urgent need to accurately identify active infection to help minimize the spread of the virus. Herein, we describe a highly specific and sensitive enzyme-linked immunosorbent assay to detect early active ZIKV using neutralizing human monoclonal antibodies isolated from infected patients in Brazil that do not cross-react with dengue viruses 1–4 and bind directly to a ZIKV immunodominant epitope. The calculated limits of detection of active ZIKV fall within the physiological ranges of the virus in human bodily fluids. This selective immunoassay creates the platform required for future translation toward a point-of-care assay for ZIKV, a necessity to diagnose active ZIKV in the remote regions of which it thrives.

Computationally Designed Peptides for Zika Virus Detection: An Incremental Construction Approach

“…The sensitivity of the three peptides decreased at the least one order of magnitude when detecting ZIKV in urine or serum. It should be highlighted that during the viremia peak of Zika infection, the lowest ZIKV concentration is 10 6.9 copies/mL in serum, close to the limit of detection showed by peptides P2 and H1 [44,45].…”

“…In all the three peptides, reactivity against ZIKV was significantly different from cross-reactivity against DENV at 10 6 copies/mL of intact virus particles, with p-values ranging from <10 −3 to <10 −4 . It is important to note that during Zika infection, the concentration of ZIKV in serum, saliva and urine can rise to around 10 6 copies/mL [44,45]. Therefore, the peptides could be suitable for discriminating between ZIKV and DENV at physiological levels.…”

Functional comparison of paper-based immunoassays based on antibodies and engineered binding proteins

“…Unfortunately, many assays have reduced function in human serum due to matrix effects and often require dilution of the serum sample in buffer to achieve reasonable performance. 7,9,12,39 To assess functionality of rcSso7d in a relevant bodily fluid, we conducted a side-by-side comparison titration with ZNS1 spiked into either PBSA or 100% human serum (Fig. 4A).…”

Advanced Nanomaterials for Preparedness Against (Re‐)Emerging Viral Diseases

“…ELISA is a standard method for screening and detecting infectious diseases in a laboratory setting. [30][31][32][33][34] It is a plate-based assay, which is available in several formats, providing results within 2-5 h. Mostly, antigens or antibodies are immobilized on the surface to form complexes with targets (such as antibodies, antigens, or whole pathogens), and specific enzyme-linked antibodies are then coupled to produce detectable signals. Unlike genetic materials, antibodies produced against a virus persist in the bloodstream following infection and vary in concentration based on the stage of infection or the period following viral exposure.…”