http://repub.eur.nl/res/aut/16030/ List of Publications en http://repub.eur.nl/static-eur/img/logo.png http://repub.eur.nl http://repub.eur.nl/res/pub/28404/ 2010-01-01T00:00:00Z Background: Rapid and specific molecular tests for identification of the recently identified pandemic influenza A/H1N1 2009 virus as well as rapid molecular tests to identify antiviral resistant strains are urgently needed. Objectives: We have evaluated the performance of two novel reverse transcriptase polymerase chain reactions (RT-PCRs) targeting specifically hemagglutinin and neuraminidase of pandemic influenza A/H1N1 virus in combination with a conserved matrix PCR. In addition, we investigated the performance of a novel discrimination RT-PCR for detection of the H275Y resistance mutation in the neuraminidase gene. Study design: Clinical performance of both subtype specific RT-PCR assays was evaluated through analysis of 684 throat swaps collected from individuals meeting the WHO case definition for the novel pandemic influenza virus. Analytical performance was analyzed through testing of 10-fold serial dilutions of RNA derived from the first Dutch sequenced and cultured confirmed case of novel pandemic influenza infection. Specificity and discriminative capacities of the H275Y discrimination assay were performed by testing wild type and recombinant H275Y pandemic influenza. Results: 121 throat swaps collected from April 2009 to July 2009 were positive by at least two out of three RT-PCRs, and negative for the seasonal H3/H1 subtype specific RT-PCR assays. 117 of these were tested positive for all three (Ct-values from 15.1 to 36.8). No oseltamivir resistance was detected. Conclusions: We present a sensitive and specific approach for detection of pandemic influenza A/H1N1 2009 and a rapid RT-PCR assay detecting a primary oseltamivir resistance mutation which can be incorporated easily into clinical virology algorithms. http://repub.eur.nl/res/pub/17683/ 2008-08-01T00:00:00Z The objective of this study was to evaluate the test characteristics of a modified BD GeneOhm methicillin-resistant Staphylococcus aureus (MRSA) assay on individual and pooled samples in a setting of low MRSA prevalence. The results of the polymerase chain reaction (PCR) assay were compared with culture results from a selective phenol red mannitol broth subcultured after 48 h. Sensitivity, specificity, and positive and negative predictive values (PPV and NPV, respectively) were calculated. For individual testing, 581 samples from 201 persons were collected; 18 (3.2%) were MRSA culture positive. Five hundred ten broths from 174 persons were combined in 106 pools after overnight incubation; 8 pools (7.5%) contained 1 or more MRSA culture-positive specimens. There were no inhibited PCR tests. The combined sensitivity of individual and pooled specimens was 92% (95% confidence interval [CI], 73-99%), the specificity was 98% (95% CI, 96-99%), and the PPV and NPV were 63% and 99.7%, respectively. Our modified procedure gives satisfactory results, and the pooling of broths may reduce costs. http://repub.eur.nl/res/pub/14012/ 2006-06-01T00:00:00Z The epidemiology and risks of Staphylococcus aureus carriage in continuous peritoneal dialysis (CPD) patients was studied in a single tertiary-care institution. On outpatient visits samples for culture were routinely taken prospectively from the CPD catheter exit site and the vestibulum nasi. Seventy-five patients with at least one culture positive for S. aureus in this period were included: 43 had genotypically identical S. aureus strains in over 80% of the cultures and were classified as persistent carriers; 32 were intermittent carriers. Persistent carriage was associated with a threefold higher risk for CPD-related infections and sixfold higher rates of vancomycin consumption compared to those for the intermittent carriers. No methicillin or vancomycin resistance was detected.