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Development of a Luciferase Immunoprecipitation Systems (LIPS) Assay to Detect IgG Antibodies against Human Respiratory Syncytial Virus Nucleoprotein

Friday, November 08, 2013 — Poster Session IV

2:00 p.m. – 4:00 p.m.

FAES Academic Center (Upper-Level Terrace)




  • S. Kumari
  • R.L. Crim
  • A. Kulkarni
  • S. Audet
  • T. Mdluli
  • H. Murata
  • J. Beeler


The nucleoprotein of respiratory syncytial virus (RSV-N) is immunogenic and elicits an IgG response following infection suggesting that anti-RSV-N antibodies may be a marker of RSV exposure. The gene for RSV-N was cloned into a mammalian expression vector, pREN2, and the expressed luciferase-tagged RSV-N protein (Ruc-N) used to develop an assay to detect anti-RSV-N specific IgG antibodies based on a high-throughput immunoprecipitation method (LIPS-N-RSV). Assay specificity was evaluated using monoclonal antibodies and pre- and post-infection/immunization serum samples from rabbits, chimpanzees, and humans with proven RSV infection. Pre-immune sera and sera from rabbits immunized with paramyxoviruses were negative when tested by LIPS-N-RSV while sera from rabbits immunized with either RSV subtype A or B gave positive signals in a dose dependent manner. An anti-RSV-N MAb was positive in the LIPS-N-RSV assay while MAbs against other myxo- and paramyxoviruses nucleoprotein and against RSV-F and -G were not. Sera from chimpanzees simultaneously immunized with vaccinia recombinant viruses expressing RSV-F and -G were negative when tested using LIPS-N-RSV, however, anti-RSV-N IgG responses were detected following live RSV challenge. LIPS-N-RSV detects RSV-N specific IgG responses that may be a useful marker of infection following RSV exposure among RSV naïve populations.

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