Characterization of Streptococcus suis serotype 2 blood infections using RT-qPCR to quantify glutamate dehydrogenase copy numbers

J Microbiol Methods. 2010 Dec;83(3):326-9. doi: 10.1016/j.mimet.2010.09.013. Epub 2010 Sep 28.

Abstract

This study characterized the dynamic distribution of bacteria in the blood of pigs infected with Streptococcus suis serotype 2 using specific primers and a TaqMan probe designed to amplify the highly conserved S. suis serotype 2 glutamate dehydrogenase (GDH) gene sequences. Gene copy numbers were used to determine the concentration of bacteria in the blood of infected pigs over time using established TaqMan real-time quantitative PCR methodologies (RT-qPCR). The results showed that the detection limit of the RT-qPCR was 10 GDH gene copies. The advantages of utilizing this approach are the high levels of specificity, sensitivity and reproducibility. Bacteria were detected in the blood of infected pigs after 24 h post infection and S. suis GDH gene copies in the experimental group were highest (10(4.15)) on day 7 post infection. Data presented in this report demonstrate that the TaqMan RT-qPCR detection method can be used to characterize the dynamic changes occurring during S. suis serotype 2 blood infections in Bama minipigs thereby facilitating research associated with defining pathogenic mechanisms associated with this organism.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics*
  • Bacterial Proteins / metabolism
  • Blood / microbiology*
  • DNA Primers / genetics
  • Gene Dosage*
  • Glutamate Dehydrogenase / genetics*
  • Glutamate Dehydrogenase / metabolism
  • Polymerase Chain Reaction / methods*
  • Streptococcal Infections / blood
  • Streptococcal Infections / diagnosis
  • Streptococcal Infections / microbiology
  • Streptococcal Infections / veterinary*
  • Streptococcus suis / enzymology
  • Streptococcus suis / genetics*
  • Swine
  • Swine Diseases / blood
  • Swine Diseases / diagnosis
  • Swine Diseases / microbiology*
  • Swine, Miniature

Substances

  • Bacterial Proteins
  • DNA Primers
  • Glutamate Dehydrogenase