title: Development of a single-tube polymerase chain reaction assay for the simultaneous detection of Haemophilus influenzae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus spp. directly in clinical samples.
abstract: This study describes the development and evaluation of a multiplex single-tube polymerase chain reaction assay for the simultaneous detection of Haemophilus influenzae, Pseudomonas aeruginosa, Staphylococcus aureus, and Streptococcus spp. used as target species-specific or genus-specific genes. The assay enables the detection of 5 to 50 pg of bacterial DNA. The sensitivity of the assay was evaluated as 100% for P. aeruginosa, S. aureus, and Streptococcus spp., and 94.3% for H. influenzae; the specificity was 100% for all 4 microorganisms (positive predictive value, 100%; negative predictive value, 98.2%). The assay permits rapid and accurate detection of these 4 microorganisms in a wide range of clinical samples such as whole blood, cerebrospinal, ear, pleural and ophthalmic fluids, as well as bronchoalveolar lavage and bronchial secretions.
authors.affiliation: National Meningitis Reference Laboratory, National School of Public Health, Athens, Greece.
authors.forename: Athanasia
authors.initials: A
authors.lastname: Xirogianni
authors.forename: Georgina
authors.initials: G
authors.lastname: Tzanakaki
authors.forename: Eleni
authors.initials: E
authors.lastname: Karagianni
authors.forename: Panayiotis
authors.initials: P
authors.lastname: Markoulatos
authors.forename: Jenny
authors.initials: J
authors.lastname: Kourea-Kremastinou
mesh_terms: D001826:Body Fluids / Q000382:microbiology; D017931:DNA Primers; D004269:DNA, Bacterial / Q000235:genetics; D006192:Haemophilus Infections / Q000382:microbiology; D006193:Haemophilus influenzae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D016920:Meningitis, Bacterial / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D013203:Staphylococcal Infections / Q000382:microbiology; D013211:Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification*; D013291:Streptococcus / Q000235:genetics / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial