title: Detection of Staphylococcus aureus enterotoxin production genes from patient samples using an automated extraction platform and multiplex real-time PCR.
abstract: To minimize specimen volume, handling and testing time, we have developed two TaqMan(®) multiplex real-time PCR (rtPCR) assays to detect staphylococcal enterotoxins A-E and Toxic Shock Syndrome Toxin production genes directly from clinical patient stool specimens utilizing a novel lysis extraction process in parallel with the Roche MagNA Pure Compact. These assays are specific, sensitive and reliable for the detection of the staphylococcal enterotoxin encoding genes and the tst1 gene from known toxin producing strains of Staphylococcus aureus. Specificity was determined by testing a total of 47 microorganism strains, including 8 previously characterized staphylococcal enterotoxin producing strains against each rtPCR target. Sensitivity for these assays range from 1 to 25 cfu per rtPCR reaction for cultured isolates and 8-20 cfu per rtPCR for the clinical stool matrix.
authors.affiliation: Wadsworth Center, New York State Department of Health, 120 New Scotland Avenue, Albany, NY, USA.
authors.forename: Amy K
authors.initials: AK
authors.lastname: Chiefari
authors.affiliation: Wadsworth Center, New York State Department of Health, 120 New Scotland Avenue, Albany, NY, USA.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Perry
authors.affiliation: Wadsworth Center, New York State Department of Health, 120 New Scotland Avenue, Albany, NY, USA.
authors.forename: Cassandra
authors.initials: C
authors.lastname: Kelly-Cirino
authors.affiliation: Wadsworth Center, New York State Department of Health, 120 New Scotland Avenue, Albany, NY, USA. Electronic address: christina.egan@health.ny.gov.
authors.forename: Christina T
authors.initials: CT
authors.lastname: Egan
mesh_terms: D057205:Automation, Laboratory; D004768:Enterotoxins / Q000235:genetics*; D005243:Feces / Q000382:microbiology; D006801:Humans; D025202:Molecular Diagnostic Techniques / Q000379:methods; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000382:microbiology; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*
chemical_list: D004768:Enterotoxins; C049463:enterotoxin E, Staphylococcal; C019368:enterotoxin D, Staphylococcal; C026066:enterotoxin A, Staphylococcal; C031818:enterotoxin B, staphylococcal; C034959:enterotoxin C, staphylococcal
keywords: Automated extraction; Diagnostic testing; Multiplex PCR; Real-time PCR; Staphylococcus aureus