title: Analysis of home dust for Staphylococcus aureus and staphylococcal enterotoxin genes using quantitative PCR.
abstract: BACKGROUND
  The bacterium Staphylococcus aureus (SA) is known to induce allergic inflammatory responses, including through secreted staphylococcal enterotoxin (SE) superantigens. To quantify indoor environmental exposures to these potential allergens, which may be associated with worse asthma, we developed a method for the assessment of S. aureus and SE in home dust and applied it to a study of homes of inner-city adults with asthma.
  
  
  METHODS
  We conducted laboratory experiments to optimize sample processing and real-time PCR methods for detection and quantification of SA (femB) and SEA-D, based on published primers. We applied this method to dust and dust extract from 24 homes. We compared results from real-time PCR to culture-based results from the same homes.
  
  
  RESULTS
  The bacteremia DNA isolation method provided higher DNA yield than alternative kits. Culture-based results from homes demonstrated 12 of 24 (50%) bedrooms were contaminated with S. aureus, only one of which carried a SE gene (SEC). In contrast, femB was detected in 23 of 24 (96%) bedrooms with a median of 1.1×106gene copies detected per gram of raw dust. Prevalence and median copy number (shown in parenthesis) of SE gene detection in bedroom dust was: SEA 25% (1.4×102); SEB 63% (1.4×103); SEC 63% (1.1×103); SED 21% (1.3×102).
  
  
  CONCLUSIONS
  Our culture-independent method to detect S. aureus and SE in home dust was more sensitive than our culture-based method. Prevalence of household exposure to S. aureus and SE allergens may be high among adults with asthma.
authors.affiliation: Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA. Electronic address: sludwig2@jhu.edu.
authors.forename: Shanna
authors.initials: S
authors.lastname: Ludwig
authors.affiliation: Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA.
authors.forename: Isabel
authors.initials: I
authors.lastname: Jimenez-Bush
authors.affiliation: Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: Emily
authors.initials: E
authors.lastname: Brigham
authors.affiliation: Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: Sonali
authors.initials: S
authors.lastname: Bose
authors.affiliation: Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: Diette
authors.affiliation: Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: Meredith C
authors.initials: MC
authors.lastname: McCormack
authors.affiliation: Johns Hopkins School of Medicine, Baltimore, MD, USA.
authors.forename: Elizabeth C
authors.initials: EC
authors.lastname: Matsui
authors.affiliation: Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, USA.
authors.forename: Meghan F
authors.initials: MF
authors.lastname: Davis
mesh_terms: D017931:DNA Primers; D004391:Dust / Q000032:analysis*; D004768:Enterotoxins / Q000032:analysis*; D005798:Genes, Bacterial; D006798:Housing; D013211:Staphylococcus aureus / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D004391:Dust; D004768:Enterotoxins
keywords: Allergy; Asthma; Indoor dust; Staphylococcal enterotoxin; Staphylococcus aureus; Superantigen