The objectives of this study were to evaluate the performance of the NucliSens easyMAG platform for nucleic acid extraction from different clinical specimens compared to NucliSens miniMAG platform and manual QIAGEN extraction. The NucliSens easyMAG and the NucliSens miniMAG showed equal performance on 215 throat swabs since real-time nucleic acid sequence-based amplification scored the same samples positive for Mycoplasma pneumoniae (n ‫؍‬ 9) and Chlamydia pneumoniae (n ‫؍‬ 5) RNAs, although internal control RNA was slightly better detected with the NucliSens easyMAG (99.3% versus 96.8%). NucliSens easyMAG extracted nucleic acids more efficiently (higher recovery and/or fewer inhibitors) compared to QIAGEN extraction by showing, on average, lower Ct values in real-time LightCycler PCR, although 4 individual specimen out of 45 were found positive only with QIAGEN. For nine M. pneumoniae-positive throat swabs, the mean difference in Ct values between NucliSens easyMAG extraction and QIAGEN extraction was ؊2.26 (range, ؊5.77 to ؉0.60); for the detection of five C. pneumoniae-positive throat swabs, the average difference in Ct values between the two methods was ؊3.38 (range, ؊6.62 to ؊2.02); and for the detection of cytomegalovirus in 24 blood samples, the mean difference in Ct values between the two methods was ؊0.95 (range, ؊5.51 to ؉1.68). The NucliSens easyMAG is considerably easier to perform, efficiently extracts nucleic acids from throat swabs and whole blood, is automated, and has high throughput.Numerous nucleic acid amplification tests are performed daily in an increasing number of clinical laboratories because of their high sensitivities and specificities. Further developments in the field have decreased the turnaround time and hands-on time. Nucleic acid extraction systems with high flexibilities in the type and number of samples to be handled and with a wide range of sample and elution volumes and short turnaround times provide a further advantage to adapt amplification techniques to clinical diagnostic requirements.A high-quality nucleic acid extract is expected to be free of amplification inhibitors and other substances that might affect enzyme substrates, and the target should be optimally recovered.The NucliSens easyMAG platform (bioMérieux, Boxtel, The Netherlands) is a second-generation system for automated isolation of nucleic acids from clinical samples based upon silica extraction technology (3). It is a benchtop instrument with the same reagents as the manual version, i.e., NucliSens miniMAG platform (bioMérieux) (4,14,17,19). Manual steps are limited to the loading of samples, reagents, and disposables. One to 24 samples can be analyzed in one run. The extraction method is universal and can be applied to a broad range of different specimens such as blood, sputum, serum, and throat swabs. The instrument can be used in combination with different amplification methods such as nucleic acid sequence-based amplification (NASBA) or PCR. Limited data are available on the NucliSens easyMAG extraction ...

BackgroundAntibiotics are frequently used in intensive care units (ICUs), and their use is associated with the emergence of bacterial resistance to antibiotics. The aim of this study was to investigate the association between the emergence of Pseudomonas aeruginosa resistance and the duration of antibiotic exposure or mode of administration in an ICU unit.MethodsA 4-year cohort study of intensive care unit was performed in patients with P. aeruginosa isolates from clinical specimens, initially susceptible to the investigated antibiotics (piperacillin/tazobactam, ceftazidime, ciprofloxacin, meropenem and amikacin). Odds ratios (ORs) with 95% confidence interval (95% CI) of emergence of resistance were calculated using logistic regression analysis for various exposure periods to antibiotics (1–3, 4–7, 8–15 and >15 days) relative to no exposure with adjustment for age, sex, Simplified Acute Physiology Score 3 (SAPS 3) and length of stay. ORs on the emergence of P. aeruginosa resistance were also calculated for the various modes of administration.ResultsIncluded were 187 patients [mean age 61 years, 69% male, mean SAPS 3 score (SD): 59 (12.3)]. None of the antibiotics investigated showed the emergence of resistance within 1–3 days. Significant meropenem resistance emerged within 8–15 days [OR 79.1 (14.9–421.0)] after antibiotic exposure unlike other antibiotics (>15 days). No difference was observed between intermittent and extended administration of meropenem and between beta-lactam mono- or combined therapy.ConclusionsUse of meropenem was associated with the emergence of resistance as soon as 8 days after exposure to the antibiotic.

Re-exposure to chickenpox may boost varicella-zoster virus (VZV) immunity in the elderly. This secondary immune response is hypothesized to confer protection against herpes zoster. We longitudinally sampled 36 adults over the course of one year after re-exposure to chickenpox. The resulting 183 samples and those of 14 controls were assessed for VZV-specific T-cell immunity and antibody titres. The percentages of VZV-specific CD4+ IL-2-producing T-cells were increased in re-exposed grandparents compared to control participants up to 9 months after re-exposure. Using a longitudinal mixture modelling approach, we found that 25% and 17% of re-exposed grandparents showed a boosting of VZV-specific CD4+ IL-2-producing T-cells and VZV-specific antibodies, respectively. The antibody boosting occurred exclusively in cytomegalovirus (CMV) IgG-positive participants. CMV IgG-positive participants also had higher VZV IE62-specific CD4+ IFN-γ-producing T-cell percentages and VZV-specific antibody titres. The protective effect of re-exposure to chickenpox is likely limited, as boosting only occurred in 17–25% of the VZV re-exposed grandparents and for less than one year.

Background Cytomegalovirus (CMV) remains an important challenge after kidney transplantation. Current Transplantation Society International Consensus Guidelines recommend antiviral prophylaxis or pre‐emptive therapy for high‐risk CMV‐seronegative recipients with a CMV‐seropositive donor (D+/R−) and moderate‐risk CMV‐seropositive recipients (R+). However, a split strategy according to CMV serostatus is not specifically mentioned. Methods We evaluated a split strategy to prevent CMV infection after kidney transplantation in which D+/R− patients received valganciclovir (VGC) prophylaxis for 200 days, and R + patients were treated pre‐emptively according to CMV DNAemia. Patients were followed until 1‐year post‐transplant. Results Between April 2014 and March 2018, 40 D+/R− and 92 R + patients underwent kidney transplantation. Forty‐six percent received antithymocyte globulin (ATG) induction, and 98% was treated with calcineurin inhibitors, mycophenolic acid (MPA), and steroids. No D+/R− patient developed CMV disease during prophylaxis (median 200 days), but 15% developed post‐prophylaxis or late‐onset disease. Fifty‐three percent developed neutropenia during prophylaxis, including 16/40 (40%) grade 3 or 4 neutropenia requiring reduction/discontinuation of MPA (30%) and/or VGC (35%), and an occasional need for granulocyte colony‐stimulating factor (5%). In the R + group, 40% received antiviral therapy for a median duration of 21 days; 5% developed early‐onset CMV disease. Only 5% developed neutropenia. D+/R + status (hazard ratio (HR) 2.09,P = .004) and ATG use (HR 2.81, P < .0001) were risk factors for CMV reactivation. Conclusions Prophylaxis leads to acceptable CMV control in high‐risk patients but comes with a high risk of neutropenia. Pre‐emptive therapy is effective and limits drug exposure in those at lower risk of CMV.