Development of an Indirect Dot-PPA-ELISA using glutamate dehydrogenase as a diagnostic antigen for the rapid and specific detection of Streptococcus suis and its application to clinical specimens

Antonie Van Leeuwenhoek. 2017 Apr;110(4):585-592. doi: 10.1007/s10482-016-0825-z. Epub 2017 Jan 5.

Abstract

Streptococcus suis is an important zoonotic pathogen causing infections in pigs and humans. Bacterial surface-related proteins are often explored as potential vaccine candidates and diagnostic antigens. In the present study, glutamate dehydrogenase, a highly conserved immunogenic extracellular protein, was used to establish a dot horseradish peroxidase enzyme-linked staphylococcal protein A immunosorbent assay (Dot-PPA-ELISA) for diagnosis of S. suis infection. The antigen-antibody reaction was optimised through checkerboard titration involving serial dilutions, followed by selective blocking tests and evaluations of cross-reaction, repeatability, and stability. Comparative analysis by using a conventional plate ELISA kit showed that the specificity and sensitivity of the Dot-PPA-ELISA were 97.5 and 96.6%, respectively. Furthermore, dynamic changes in the levels of antibody in rabbits immunised with a propolis inactivated vaccine were monitored by Dot-PPA-ELISA. A total seroprevalence of 73.1% in 305 pig serum samples indicated the method's applicability to detect S. suis infection. Cumulatively, the results suggested that Dot-PAA-ELISA is a convenient, rapid, sensitive, and specific diagnostic method suitable for studying large numbers of samples obtained from clinical and epidemiological studies, thereby helping reduce important economic losses.

Keywords: Diagnosis; Dot-PPA-ELISA; Streptococcus suis; rGDH.

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood*
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Glutamate Dehydrogenase / immunology*
  • Rabbits
  • Sensitivity and Specificity
  • Streptococcal Infections / diagnosis*
  • Streptococcal Infections / microbiology
  • Streptococcal Infections / veterinary*
  • Streptococcus suis / immunology*

Substances

  • Antibodies, Bacterial
  • Glutamate Dehydrogenase