We evaluated the Prodesse ProFlu-1 real-time reverse transcription-PCR multiplex assay with the SmartCycler instrument for the detection of human respiratory syncytial virus (RSV) and influenza A and B viruses in comparison to conventional cell culture and antigen immunoassays with the BD Directigen A؉B and Binax NOW RSV assays over two successive respiratory virus seasons. Ninety-two percent of the 361 specimens tested were nasopharyngeal aspirates obtained from individual patients, of which 119 were positive for RSV and 59 were positive for influenza virus. The median age of the patients whose specimens were positive for RSV and influenza virus were 6.3 months and 42.4 years, respectively. The specificity of all of the methods tested was >99%, and the individual sensitivities of NOW RSV, RSV culture, Directigen A؉B, influenza virus culture, and the Proflu-1 PCR for influenza/RSV were 82% (95% confidence interval [CI], 73 to 88), 57% (95% CI, 44 to 69), 59% (95% CI, 44 to 72), 54% (95% CI, 38 to 69), and 98% (95% CI, 93 to 100)/95% (95% CI, 85 to 99), respectively. In a clinical setting where viral isolation is performed to confirm rapid antigen immunoassay results for these common respiratory viruses, one-step real-time reverse transcriptase PCR testing can be a more sensitive and timely confirmatory method.Human respiratory syncytial virus (RSV) and influenza A and B viruses are respiratory pathogens associated with substantial morbidity and mortality annually (43). Virtually all children become infected with RSV within 2 years after birth, and 1% require hospitalization (15). Although the importance of RSV as a cause of pneumonia and brochiolitis in young children is well recognized (21), the most serious morbidity and highest mortality associated with both RSV and influenza virus circulation occurs disproportionately among elderly persons (43). The first-line tests used to detect these virus infections in many hospitals are antigen-based immunoassays. It has been demonstrated that antigen immunoassays have exceedingly poor sensitivity in detecting RSV and influenza virus infections in the elderly, seriously limiting their utility for detecting and confirming institutional or community outbreaks (7,13,38). This study was intended to evaluate the performance of viral isolation in cell culture, one-step real-time multiplex reverse transcription-PCR (RT-PCR), and antigen immunoassays for the detection of influenza virus and RSV in respiratory specimens from adults and children during two respiratory virus seasons. MATERIALS AND METHODS Specimens.Upper respiratory tract specimens were collected from 353 individual symptomatic patients during two successive winter respiratory virus seasons encompassing

Emerging contaminants, including pharmaceutical and personal care products, are receiving considerable attention owing to their potential to negatively impact the environment and to pose risks to human health. The widespread use of antibiotics and their fate and transport in the environment pose further risks with respect to public health and the development of antibiotic resistant organisms (AROs). While the occurrence of AROs is important, there is increasing interest in antibiotic resistance genes (ARGs). An urgent need exists to improve our understanding of the mechanisms associated with the spread and development of ARGs in both clinical and veterinary settings, the human body (gastrointestinal tract and microbiome) as well as in engineered (wastewater treatment plants) and natural (soil, sediments and water) environments. This review focuses on ARGs as an emerging environmental contaminant. The factors and mechanisms involved in ARG dissemination in a variety of environments are explored in detail. The unique challenges of ARGs with respect to policy-making and environmental monitoring are identified, and recommendations regarding how these challenges might be addressed are provided.

The MRSA-Screen (Denka-Seiken, Tokyo, Japan) latex agglutination test was evaluated for its ability to detect PBP 2a from 200 clinical isolates of coagulase-negative staphylococci (CoNS; 84 mecA-positive strains and 116 mecA-negative strains) consisting of 108 Staphylococcus epidermidis, 37 S. saprophyticus, 15 S. haemolyticus, 11 S. hominis, 10 S. capitis, 10 S. warneri, and 3 S. lugdunensis species as well as 6 other species of CoNS. The assay was compared with susceptibility testing with an agar screen plate with oxacillin at 6 g/ml (OXA6), by oxacillin disk diffusion (DD), by broth microdilution (BMDIL), by the E test, and with Vitek GPS-SV and Vitek GPS-107 susceptibility cards. PCR for the detection of the mecA gene was used as the "gold standard." The sensitivities and specificities for the methods evaluated were as follows: MRSA-Screen, 100 and 100%, respectively; OXA6, 100 and 99%, respectively; DD, 98 and 62%, respectively; BMDIL, 100 and 60%, respectively; E test, 100 and 51%, respectively; Vitek GPS-SV susceptibility card, 98 and 87%, respectively; and Vitek GPS-107 susceptibility card, 100 and 61%, respectively. The MRSA-Screen test accurately and rapidly detected oxacillin resistance in CoNS.Coagulase-negative staphylococci (CoNS) are recognized as important causes of nosocomial infection, especially in neonates, those who are immunocompromised, and patients with indwelling prosthetic devices (6). The rate of resistance to oxacillin among CoNS has been increasing (2), and the empirical treatment of choice for infections caused by these organisms is often vancomycin. The rapid and accurate identification of oxacillin resistance is essential in order to determine the most appropriate antimicrobial therapy. The major mechanism of oxacillin resistance in staphylococci is mediated by the production of a modified penicillin-binding protein, PBP 2a, specified by the mecA gene. In 1999, the National Committee for Clinical Laboratory Standards (NCCLS) lowered the breakpoint for determination of oxacillin resistance in CoNS from Ն4 to Ն0.5 g/ml (12) in order to more accurately reflect mecA-mediated resistance. In addition, the use of the oxacillin agar screen plate (Mueller-Hinton agar with 6 g of oxacillin per ml supplemented with 4% NaCl) was no longer recommended by NCCLS for the detection of oxacillin resistance in CoNS, as reports in the literature had indicated that this method may not be able to detect mecA-mediated oxacillin resistance in these strains (15,17).A rapid latex agglutination test, the MRSA-Screen (DenkaSeiken, Tokyo, Japan), has recently been developed in order to detect the presence of oxacillin resistance mediated by PBP 2a. Although several studies have evaluated the use of this test for the detection of oxacillin resistance in Staphylococcus aureus (8,18,19), its accuracy for the detection of oxacillin resistance in CoNS has not been extensively assessed (5, 9). In the study described here, we evaluated the MRSA-Screen for its ability to detect PBP 2a in clinical strains of CoNS. In ...