Rapid and Sensitive Detection of Venezuelan Equine Encephalitis Virus by Electrochemiluminescence Immunoassay

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Authors
  1. Dai, X.
  2. Fulton, R.E.
  3. Hilsen, R.
  4. Hu, W.G.
  5. Ranches, J.
Corporate Authors
Defence R&D Canada - Suffield, Ralston ALTA (CAN)
Abstract
A sensitive and rapid electrochemiluminescence (ECL) immunoassay to detect and identify Venezuelan equine encephalitis virus (VEEV) was developed using the BioVeris M1MR system. VEEV strain TC-83 and recombinant VEEV envelope E2 protein were used as target antigens in the ECL assay. Under optimized conditions, the assay limit of detection (LOD) was 103 pfu/mL for TC-83 whole virus and 1 ng/mL for recombinant E2. Comparison of three antibody pairs against the different antigen targets demonstrated that some antibody pairs maintained high reactivity with both whole virus and recombinant E2, while others had reactivity with either whole virus or recombinant E2. Testing of samples over time (month to month) and by different operators (person to person) indicated that the assay was reproducible with the coefficient of variation ranging from 4.7% to 18.5%. In addition, a genetically biotinylated recombinant antibody, MA116SBP, was tested and the assay results were compared to those obtained using chemically biotinylated antibodies. Results indicated that MA116SBP was functional in the ECL assay but resulted in a higher LOD to that obtained using chemically biotinylated antibodies (100 ng/mL vs 1 ng/mL). The ECL assay was specific to VEEV as evidenced by the absence of crossreactivity with two closely related alphaviruses and twenty-one other bacterial, viral, and toxin agents. The ECL assay was found to be better in terms of sensitivity, assay time, and automation than the standard

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Keywords
Electrochemiluminescence immunoassay;ECL;M1MR;Venezuelan equine encephalitis virus;Recombinant E2 protein;VEE virus
Report Number
DRDC-SUFFIELD-TR-2008-050 — Technical Report
Date of publication
01 Feb 2008
Number of Pages
42
DSTKIM No
CA030776
CANDIS No
529419
Format(s):
CD ROM

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