The Use of NanoTrap Particles as a Sample Enrichment Method to Enhance the Detection of Rift Valley Fever Virus

Autor: Alan Baer, Chelsea Pinkham, Fatah Kashanchi, Charles L. Bailey, Kylene Kehn-Hall, Aarthi Narayanan, Katherine Fite, Benjamin Lepene, Nazly Shafagati
Rok vydání: 2013
Předmět:
Zdroj: PLoS Neglected Tropical Diseases, Vol 7, Iss 7, p e2296 (2013)
PLoS Neglected Tropical Diseases
ISSN: 1935-2735
DOI: 10.1371/journal.pntd.0002296
Popis: Background Rift Valley Fever Virus (RVFV) is a zoonotic virus that is not only an emerging pathogen but is also considered a biodefense pathogen due to the threat it may cause to public health and national security. The current state of diagnosis has led to misdiagnosis early on in infection. Here we describe the use of a novel sample preparation technology, NanoTrap particles, to enhance the detection of RVFV. Previous studies demonstrated that NanoTrap particles lead to both 100 percent capture of protein analytes as well as an improvement of more than 100-fold in sensitivity compared to existing methods. Here we extend these findings by demonstrating the capture and enrichment of viruses. Results Screening of NanoTrap particles indicated that one particle, NT53, was the most efficient at RVFV capture as demonstrated by both qRT-PCR and plaque assays. Importantly, NT53 capture of RVFV resulted in greater than 100-fold enrichment from low viral titers when other diagnostics assays may produce false negatives. NT53 was also capable of capturing and enhancing RVFV detection from serum samples. RVFV that was inactivated through either detergent or heat treatment was still found bound to NT53, indicating the ability to use NanoTrap particles for viral capture prior to transport to a BSL-2 environment. Furthermore, both NP-40-lysed virus and purified RVFV RNA were bound by NT53. Importantly, NT53 protected viral RNA from RNase A degradation, which was not observed with other commercially available beads. Incubation of RVFV samples with NT53 also resulted in increased viral stability as demonstrated through preservation of infectivity at elevated temperatures. Finally, NanoTrap particles were capable of capturing VEEV and HIV, demonstrating the broad applicability of NanoTrap particles for viral diagnostics. Conclusion This study demonstrates NanoTrap particles are capable of capturing, enriching, and protecting RVFV virions. Furthermore, the use of NanoTrap particles can be extended to a variety of viruses, including VEEV and HIV.
Author Summary There is a dire need for fast and efficient diagnosis of many viral diseases. Our research specifically looked at RVFV, a virus that can only be worked with in biosafety level 3 (BSL-3) laboratories, and its capture with NanoTrap particles. NanoTrap particles are hydrogel particles that contain internal affinity baits. They have previously been used in the capture of several analytes, but never in the capture of whole virus particles. We were not only able to capture and detect RVFV at very low titers from both media and serum, but we were also able to inactivate the virus, which allows for its safe transport to BSL-2 laboratories. While there are other commercially available beads that can also capture virus, NanoTrap particles are the only beads that can protect the viral RNA from enzymatic degradation. Furthermore, we demonstrated that whole virus detection with NanoTrap particles is not limited to only RVFV, but that NanoTrap particles can be used to detect other viruses such as Human Immunodeficiency Virus (HIV) and Venezuelan Equine Encephalitis Virus (VEEV).
Databáze: OpenAIRE
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