Microbead electrochemiluminescence immunoassay for detection and identification of Venezuelan equine encephalitis virus.
Summary of "Microbead electrochemiluminescence immunoassay for detection and identification of Venezuelan equine encephalitis virus."
An electrochemiluminescence (ECL) immunoassay, incorporating chemically biotinylated and ruthenylated antibodies down-selected from a panel of monoclonal and polyclonal reagents, was developed to detect and identify Venezuelan equine encephalitis virus (VEEV). The limit of detection (LOD) of the optimized ECL assay was 10(3) pfu/ml VEEV TC-83 virus and 1ng/ml recombinant (r) VEEV E2 protein. The LOD of the ECL assay was approximately one log unit lower than that of a sandwich enzyme-linked immunosorbent assay (ELISA) incorporating the same immunoreagents. Repetition of ECL assays over time and by different operators demonstrated that the assay was reproducible (coefficient of variation 4.7-18.5% month-to-month; 3.3-8.8% person-to-person). The VEEV ECL assay exhibited no cross-reactivity with two closely related alphaviruses or with 21 heterologous biological agents. A genetically biotinylated recombinant VEEV antibody, MA116SBP, was evaluated for utility for detection of rE2; although functional in the ECL assay, the LOD was two log units higher (100ng/ml vs 1ng/ml) using MA116SBP than when chemically biotinylated antibody was used. The ECL assay detected VEEV at the lowest LOD (highest sensitivity) hitherto reported in the published literature and ECL assay results were generated in approximately 60min compared to a 6-8 hr period required for ELISA. Results have demonstrated a sensitive, rapid, and fully automated ECL immunoassay for detection and identification of VEEV.
Affiliation
Defence Research and Development Canada-Suffield, PO Box 4000, Station Main, Medicine Hat, AB T1A 8K6.
Journal Details
This article was published in the following journal.
Name: Journal of virological methods
ISSN: 1879-0984
Pages:
Links
- PubMed Source: http://www.ncbi.nlm.nih.gov/pubmed/20678522
- DOI: http://dx.doi.org/10.1016/j.jviromet.2010.07.022
Medical and Biotech [MESH] Definitions
Encephalomyelitis, Venezuelan Equine
A form of arboviral encephalitis endemic to Central America and the northern latitudes of South America. The causative organism (ENCEPHALITIS VIRUS, VENEZUELAN EQUINE) is transmitted to humans and horses via the bite of several mosquito species. Human viral infection may be asymptomatic or remain restricted to a mild influenza-like illness. Encephalitis, usually not severe, occurs in a small percentage of cases and may rarely feature SEIZURES and COMA. (From Joynt, Clinical Neurology, 1996, Ch26, pp9-10)
Encephalitis Virus, Venezuelan Equine
A species of ALPHAVIRUS that is the etiologic agent of encephalomyelitis in humans and equines. It is seen most commonly in parts of Central and South America.
Encephalomyelitis, Eastern Equine
A form of arboviral encephalitis (primarily affecting equines) endemic to eastern regions of North America. The causative organism (ENCEPHALOMYELITIS VIRUS, EASTERN EQUINE) may be transmitted to humans via the bite of AEDES mosquitoes. Clinical manifestations include the acute onset of fever, HEADACHE, altered mentation, and SEIZURES followed by coma. The condition is fatal in up to 50% of cases. Recovery may be marked by residual neurologic deficits and EPILEPSY. (From Joynt, Clinical Neurology, 1996, Ch26, pp9-10)
Encephalitis Viruses, Japanese
A subgroup of the genus FLAVIVIRUS which comprises a number of viral species that are the etiologic agents of human encephalitis in many different geographical regions. These include Japanese encephalitis virus (ENCEPHALITIS VIRUS, JAPANESE), St. Louis encephalitis virus (ENCEPHALITIS VIRUS, ST. LOUIS), Murray Valley encephalitis virus (ENCEPHALITIS VIRUS, MURRAY VALLEY), and WEST NILE VIRUS.
Gonadotropins, Equine
Gonadotropins secreted by the pituitary or the placenta in horses. This term generally refers to the gonadotropins found in the pregnant mare serum, a rich source of equine CHORIONIC GONADOTROPIN; LUTEINIZING HORMONE; and FOLLICLE STIMULATING HORMONE. Unlike that in humans, the equine LUTEINIZING HORMONE, BETA SUBUNIT is identical to the equine choronic gonadotropin, beta. Equine gonadotropins prepared from pregnant mare serum are used in reproductive studies.
PubMed Articles
The requirement to replicate in both vertebrate and invertebrate hosts is thought to limit the introduction of genetic changes into the genome of arboviruses. Serial passage under laboratory condition...
TC83 is a human vaccine with investigational new drug status and is used as a prototype Venezuelan equine encephalitis virus for pathogenesis and antiviral research. Differing from other experimental...
The present study identified a linear B-cell epitope in the Eastern equine encephalitis virus (EEEV) E2 glycoprotein by screening a phage-displayed random 12-mer peptide library using an EEEV E2 speci...
We have previously shown that a hydrophobic alkylating compound, 1,5-iodonaphthyl-azide (INA) can efficiently inactivate the virulent strain of Venezuelan equine encephalitis virus (VEEV), V3000 in vi...
We recently described a Venezuelan equine encephalitis virus (VEEV)-specific human monoclonal antibody (MAb), F5 nIgG, that recognizes a new neutralization epitope on the VEEV E2 envelope glycoprotein...
Clinical Trials
This study is being done to determine the safety and tolerability of a new investigational vaccine referred to as VEE IA/B V3526, which may induce production of specific antibodies in vacc...
Safety and Immunogenicity Study of the Venezuelan Equine Encephalomyelitis Vaccine
This study is designed to determine safety of and immune response to Venezuelan Equine Encephalomyelitis Vaccine, Live, Attenuated, Dried TC-83, NDBR-102 (TC-83).
Western Equine Encephalitis Vaccine
This study is designed to determine the safety and immunogenicity of WEE Vaccine Lot number 3-1-92.
Safety and Immunogenicity Study of Eastern Equine Encephalitis (EEE) Vaccine
This study is designed to determine the safety and immunogenicity of Eastern Equine Encephalitis (EEE) Vaccine
Study of ChimeriVax™-Japanese Encephalitis Vaccine in Toddlers in Thailand and the Philippines
This study is to assess the safety and immunogenicity of three consecutive lots of JE-CV in toddlers aged 12-18 months. Primary objective: To demonstrate the bio-equivalence of thr...
