Integrating Magnetic-Bead-Based Sample Extraction and Molecular Barcoding for the One-Step Pooled RT-qPCR Assay of Viral Pathogens without Retesting.

Pooling multiple samples prior to real-time reverse-transcription polymerase chain reaction (RT-PCR) analysis has been proposed as a strategy to minimize expenses and boost test throughput during the COVID-19 pandemic. Nevertheless, the traditional pooling approach cannot be effectively deployed in high-prevalence settings due to the need for secondary tests in the case of a positive pool. In this study, we present a pooling test platform with high adaptability and simplicity that allows sample-specific detection of multiple-tagged samples in a single run without the need for retesting. This was accomplished by labeling distinct samples with predefined ID-Primers and identifying tagged pooled samples using one-step RT-PCR followed by melting curve analysis with rationally designed universal fluorescence- and quencher-tagged oligo probes. Using magnetic beads (MBs), nucleic acid targets from different individuals can be tagged and extracted concurrently and then pooled before RT, eliminating the need for extra RNA extraction and separate RT and enzyme digestion steps in the recently developed barcoding strategies. Pools of six samples (positive and negative) were successfully identified by melting temperature values under two fluorescent channels, with a detection sensitivity of 5 copies/µL. We validated the reproducibility of this assay by running it on 40 clinical samples with a hypothetical infection rate of 15%. In addition, to aid the scenario of large-scale pooling tests, we constructed a melting curve autoreadout system (MCARS) for statistical analysis of melting curve plots to eliminate error-prone manual result readout. Our results suggest that this strategy could be a simple and adaptable tool for alleviating existing bottlenecks in diagnostic pooling testing.

Wound complications after open reduction and internal fixation of tibial plateau fractures in the elderly: a multicentre study.

PURPOSE: The incidence of wound complications after open reduction with internal fixation (ORIF) of tibial plateau fractures in young patients has been reported to range from approximately 5 to 15%. Reports on wound complication rates in the elderly patients are limited. This study investigates the incidence of post-operative wound complications in elderly patients undergoing ORIF of their tibial plateau fractures. METHODS: A retrospective study was performed within three accredited level 1 trauma centres. Patients > 60 years of age undergoing open reduction and internal fixation of their tibial plateau fractures were included. The primary outcome measure was wound complications of the surgical site. These were divided into superficial infections versus deep infections. RESULTS: One hundred two patients matched the inclusion criteria. Of these, 16 patients (15.7%) developed a post-operative wound infection. The analysis of underlying co-morbidities and risk factors revealed that patients with American Society of Anaesthesiologists (ASA) classes 3 and 4 were at significantly increased risk of sustaining a wound complications as compared to ASA classes 1 and 2 (23.7 versus 5.1%, p = 0.015). CONCLUSIONS: The overall infection rates in elderly patients undergoing ORIF for tibial plateau fractures is in a similar range to published data on younger patient populations. In particular, elderly patients without significant co-morbidities seem to be appropriate candidates for ORIF of their tibial plateau fractures. However, elderly patients with significant co-morbidities must be considered as high risk and alternative treatment options, such as nonoperative treatment or less invasive surgical options, should be explored in these patients.