Molecular Detection of Filamentous Fungi in Formalin-Fixed Paraffin-Embedded Specimens in Invasive Fungal Wound Infections Is Feasible with High Specificity.

Trauma-related invasive fungal wound infections (IFIs) are associated with significant morbidity and mortality. Early identification and treatment are critical. Traditional identification methods (e.g., fungal cultures and histopathology) can be delayed and insensitive. We assessed a PCR-based sequencing assay for rapid identification of filamentous fungi in formalin-fixed paraffin-embedded (FFPE) specimens obtained from combat casualties injured in Afghanistan. Blinded FFPE specimens from cases (specimens positive on histopathology) and controls (specimens negative on histopathology) were submitted for evaluation with a panfungal PCR. The internal transcribed spacer 2 (ITS2) region of the fungal ribosomal repeat was amplified and sequenced. The PCR results were compared with findings from histopathology and/or culture. If injury sites contributed multiple specimens, findings for the site were collapsed to the site level. We included 64 case subjects (contributing 95 sites) and 102 controls (contributing 118 sites). Compared to histopathology, panfungal PCR was specific (99%), but not as sensitive (63%); however, sensitivity improved to 83% in specimens from sites with angioinvasion. Panfungal PCR identified fungi of the order Mucorales in 33 of 44 sites with angioinvasion (75%), whereas fungal culture was positive in 20 of 44 sites (45%). Saksenaea spp. were the dominant fungi identified by PCR in specimens from angioinvasion sites (57%). Panfungal PCR is specific, albeit with lower sensitivity, and performs better at identifying fungi of the order Mucorales than culture. DNA sequencing offers significant promise for the rapid identification of fungal infection in trauma-related injuries, leading to more timely and accurate diagnoses.

Practical Use of Metal Oxide Semiconductor Gas Sensors for Measuring Nitrogen Dioxide and Ozone in Urban Environments.

The potential of inexpensive Metal Oxide Semiconductor (MOS) gas sensors to be used for urban air quality monitoring has been the topic of increasing interest in the last decade. This paper discusses some of the lessons of three years of experience working with such sensors on a novel instrument platform (Small Open General purpose Sensor (SOGS)) in the measurement of atmospheric nitrogen dioxide and ozone concentrations. Analytic methods for increasing long-term accuracy of measurements are discussed, which permit nitrogen dioxide measurements with 95% confidence intervals of 20.0 µ g m - 3 and ozone precision of 26.8 µ g m - 3 , for measurements over a period one month away from calibration, averaged over 18 months of such calibrations. Beyond four months from calibration, sensor drift becomes significant, and accuracy is significantly reduced. Successful calibration schemes are discussed with the use of controlled artificial atmospheres complementing deployment on a reference weather station exposed to the elements. Manufacturing variation in the attributes of individual sensors are examined, an experiment possible due to the instrument being equipped with pairs of sensors of the same kind. Good repeatability (better than 0.7 correlation) between individual sensor elements is shown. The results from sensors that used fans to push air past an internal sensor element are compared with mounting the sensors on the outside of the enclosure, the latter design increasing effective integration time to more than a day. Finally, possible paths forward are suggested for improving the reliability of this promising sensor technology for measuring pollution in an urban environment.