Abstract:
Objective Lymphocytic choriomeningitis virus (LCMV) is an important pathogen of human meningitis and a globally distributed yet long-neglected zoonotic virus. To investigate LCMV infection in population in China, we established a luciferase immunoprecipitation system (LIPS) based on the LCMV nucleoprotein (NP) for the detection of LCMV NP-specific antibody in human serum samples.
Methods A eukaryotic expression plasmid encoding an NP-luciferase fusion protein was constructed and transfected into eukaryotic cells. Cell lysates containing expressed luciferase-labeled nucleoprotein antigen (Luc-NP) solution were collected for the assay development. By using LCMV-positive and LCMV-negative rabbit sera, a preliminary LIPS-based serological assay was established and the reaction conditions were further optimized, including incubation temperature, amount of recombinant, and agarose volume. The established LIPS assay was then used to detect NP-specific antibody in 200 serum samples collected from individuals undergoing routine health examinations in Lingao county, Hainan province.
Results The Luc-NP antigen was successfully constructed and expressed, which was confirmed by indirect immunofluorescence assay and western blotting. The optimal LIPS conditions were determined to be incubation at room temperature with 20 μL of agarose in the reaction system. The LIPS assay was able to discriminate LCMV from plateau pika virus, but not LCMV from Wenzhou virus. In the 200 human serum samples tested, one sample was positive (0.50%), the results was consistent with those obtained by indirect immunofluorescence assay and western blotting.
Conclusion A preliminary LIPS assay based on the LCMV NP protein was established and successfully applied in the detection of LCMV antibody in human serum samples.
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