Latex agglutination test for rapid on-site serodiagnosis of Japanese encephalitis in pigs using recombinant NS1 antigen

J Vector Borne Dis. 2019 Apr-Jun;56(2):105-110. doi: 10.4103/0972-9062.263717.

Abstract

Background & objectives: Japanese encephalitis (JE) is a mosquitoe-borne viral zoonotic disease and globally around three billion people are at the risk of disease. The occurrence of JE cases has shown a rising trend during last decade in India. Pig is the amplifying host for JE virus and serves as a suitable sentinel model for the prediction of disease outbreak in humans. The development of a diagnostic test that is suitable for surveillance of JE in pigs is the need of the hour. The existing tests require elaborate laboratory facilities which make their application in rural settings difficult. Therefore, realizing the need for a rapid test, efforts were made to standardize a latex agglutination test (LAT) for serodiagnosis of JE in pigs.

Methods: Standardization of LAT by physical adsorption of recombinant NS1 (non-structural) protein of JE virus onto latex beads was done by altering six different variables, namely the antigen concentration, sensitization condition, surface blocking agent, blocking condition, particle concentration and reaction time. The standardized latex-protein complex was used for screening 246 pig serum samples under optimal conditions.

Results: The test was standardized with a diagnostic sensitivity and specificity of 82.24 and 87.83%, respectively. Screening of 246 field pig serum samples using standardized LAT showed a seropositivity of 50.4%. The results were available within 5 min after addition of test serum sample to the sensitized beads.

Interpretation & conclusion: The findings of the study highlight the potential of LAT as a rapid on-site assay for JE diagnosis in pigs which would aid in predicting JE outbreaks in humans.

Keywords: Japanese encephalitis; NS1 antigen; latex agglutination test; serodiagnosis.

MeSH terms

  • Animals
  • Antibodies, Viral / blood*
  • Encephalitis Virus, Japanese / immunology
  • Encephalitis, Japanese / immunology*
  • India
  • Latex Fixation Tests / standards*
  • Latex Fixation Tests / veterinary
  • Recombinant Proteins / immunology
  • Sensitivity and Specificity
  • Swine
  • Viral Nonstructural Proteins / genetics*
  • Viral Nonstructural Proteins / immunology
  • Zoonoses / diagnosis*
  • Zoonoses / immunology
  • Zoonoses / virology

Substances

  • Antibodies, Viral
  • NS1 protein, Flavivirus
  • Recombinant Proteins
  • Viral Nonstructural Proteins