Abstract
Most of the modern techniques used for identification of viral-induced disease are based on identification of viral antigens and/or nucleic acids in patient's blood. Diagnosis in the field or in remote locations can be challenging and alternatively samples are shipped to diagnostic labs for testing. Shipments must occur under controlled temperature conditions to prevent loss of sample integrity. We have tested the ability of magnetic Nanotrap® (NT) particles to improve stability and detection of Venezuelan equine encephalitis virus (VEEV), viral capsid protein, and viral genomic RNA in whole human blood at elevated temperature and prolonged storage conditions. NT particles have previously been shown to capture and enrich multiple pathogens including respiratory syncytial virus, influenza virus, coronavirus, and Rift Valley fever virus. Our study indicates that samples incubated with NT particles had detectable levels of infectious VEEV in blood equal to or greater than samples without NT treatment across all temperatures. Viral RNA detection was increased in the presence of NT particles at later time points (72 h) and higher temperature (40°C) conditions. Likewise, detection of VEEV capsid protein was enhanced in the presence of NT particles up to 72 h at 40°C. Finally, we intranasally infected C3H mice with TC-83, the live attenuated vaccine strain of VEEV, and demonstrated that NT particles could substantially increase the detection of VEEV capsid in infected blood incubated up to 72 h at 40°C. Samples without NT particles had undetectable capsid protein levels. Taken together, our data demonstrate the ability of NT particles to preserve and enable detection of VEEV in human and mouse blood samples over time and at elevated temperatures.
Highlights
Venezuelan equine encephalitis virus (VEEV) is one of the most neglected viruses among biowarfare agents
Our results indicate that: (i) magnetic NT particles enhance preservation of infectious VEEV in whole human blood at 40◦C; (ii) NT maintain significantly higher levels of VEEV RNA in whole human blood at 40◦C; (iii) NT retain their functional activity at both normal and elevated humidity conditions and significantly preserve VEEV infectivity in such an environment; and (iv) blood samples from VEEV TC-83 infected animals are better protected from capsid protein degradation if they are incubated with NT
Our results demonstrate for the first time the capability of NT particles to stabilize virus in blood at elevated temperatures, the direct interaction of NT particles with VEEV, and the utility of NT particles with viral clinical samples
Summary
Venezuelan equine encephalitis virus (VEEV) is one of the most neglected viruses among biowarfare agents It is classified as a Category B biothreat pathogen by National Institute of Allergy and Infectious Diseases (NIAID), USA, due to its high dissemination rate, minimal infectious dose to induce disease in human, and the requirement for specific and enhanced diagnostic capacities. VEEV causes disease with symptoms ranging from influenzalike illness to more severe illnesses including myalgia, arthralgia, and neurological disorders that can lead to lethal encephalitis in susceptible hosts. All these characteristics made VEEV an attractive candidate for weaponization [1]. The spillover of VEEV infection from infected horses to humans during epidemics remains a concern
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