Identification and Characterization of Staphylococcus aureus Strains with an Incomplete Hemolytic Phenotype

Front Cell Infect Microbiol. 2016 Nov 18:6:146. doi: 10.3389/fcimb.2016.00146. eCollection 2016.

Abstract

Staphylococcus aureus is a common pathogen causing both hospital and community-acquired infections. Hemolysin is one of the important virulence factors for S. aureus and causes the typical β-hemolytic phenotype which is called complete hemolytic phenotype as well. Recently, S. aureus with an incomplete hemolytic phenotype (SIHP) was isolated from clinical samples. To study the microbiologic characteristics of SIHP, the special hemolytic phenotype of SIHP was verified on the sheep blood agar plates supplied by different manufacturers. Expression of hemolysin genes hla, hlb, hlgC, and hld of SIHP was detected by qRT-PCR and it was showed that expression of hlb in SIHP was obviously increased compared to the control S. aureus strains with complete hemolytic phenotype (SCHP), while the expression of hla, hlgC, and hld in SIHP was significantly decreased. In addition, the α-hemolysin encoded by gene hla was decreased obviously in SIHP compared to SCHP by western blot. All 60 SIHP strains were identified to be the methicillin resistant S. aureus (MRSA), and moreover these SIHP strains all contains mecA gene. The virulence gene tst were all present in SIHP, and the intracellular survival ability of SIHP was much greater than that of the gene tst negative S. aureus. We also found that IL-2, IL-6, and IL-17A secreted in the supernatant of SIHP infected macrophages increased significantly compared to tst negative control strains infected ones. MLST analysis showed that all of SIHP strains were classified into ST5 clone. To our knowledge, this study firstly showed that SIHP strains are a kind of methicillin resistant strains which express β-hemolysin highly and possess a potential high virulence, and it was suggested that SIHP should be paid more attention in hospital.

Keywords: Staphylococcus aureus; drug resistance; hemolysin; incomplete hemolytic phenotype; virulence.

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Toxins / genetics*
  • Cell Line
  • Chemokines / metabolism
  • Cytokines / metabolism
  • Enterotoxins / genetics
  • Extracellular Matrix Proteins / genetics
  • Gene Expression Regulation, Bacterial
  • Genes, Bacterial
  • Hemolysin Proteins / genetics*
  • Humans
  • Interleukin-17 / metabolism
  • Interleukin-2 / metabolism
  • Interleukin-6 / metabolism
  • Macrophages / metabolism
  • Macrophages / microbiology
  • Methicillin-Resistant Staphylococcus aureus
  • Microbial Sensitivity Tests
  • Multilocus Sequence Typing
  • Penicillin-Binding Proteins / genetics
  • Phenotype*
  • RNA, Messenger / biosynthesis
  • Sheep
  • Sphingomyelin Phosphodiesterase / genetics*
  • Staphylococcal Infections / microbiology
  • Staphylococcus aureus / classification
  • Staphylococcus aureus / genetics*
  • Staphylococcus aureus / isolation & purification*
  • Superantigens / genetics
  • Survival
  • Virulence Factors / genetics*

Substances

  • Bacterial Proteins
  • Bacterial Toxins
  • Chemokines
  • Cytokines
  • Enterotoxins
  • Extracellular Matrix Proteins
  • Hemolysin Proteins
  • Interleukin-17
  • Interleukin-2
  • Interleukin-6
  • Penicillin-Binding Proteins
  • RNA, Messenger
  • Superantigens
  • Virulence Factors
  • enterotoxin F, Staphylococcal
  • mecA protein, Staphylococcus aureus
  • staphylococcal alpha-toxin
  • Sphingomyelin Phosphodiesterase
  • hlb protein, Staphylococcus aureus