RESUMO
BackgroundSurveillance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infections is essential for the global containment measures with regard to the ongoing pandemic. Diagnostic gold standard is currently reverse transcription of the (+)RNA genome and subgenomic RNAs and subsequent quantitative polymerase chain reaction (RT-qPCR) from nasopharyngeal swabs or bronchoalveolar lavages. In order to further improve the diagnostic accuracy, particularly for the reliable discrimination between negative and false-negative specimens, we propose the combination of the RT-qPCR workflow with subsequent pyrosequencing of a S-gene amplicon. This extension might add important value mainly in cases with low SARS-CoV-2 load, where RT-qPCR alone can deliver conflicting results.s ResultsWe successfully established a combined RT-qPCR and S-gene pyrosequencing method which can be optionally exploited after routine diagnostics or for epidemiologic studies. This allows a more reliable interpretation of conflicting RT-qPCR results in specimens with relatively low viral loads and close to the detection limits of qPCR (CT values >30). After laboratory implementation and characterization of a best practice protocol we tested the combined method in a large pediatric cohort from two German medical centers (n=769). Pyrosequencing after RT-qPCR enabled us to uncover 6 previously unrecognized cases of pediatric SARS-CoV-2 associated diseases, partially exhibiting unusual and heterogeneous presentation. Moreover, it is notable that in the course of RT-qPCR/pyrosequencing method establishment we did not observe any case of false-positive diagnosis when confirmed SARS-CoV-2-positive specimens were used from foregoing routine testing. ConclusionsThe proposed protocol allows a specific and sensitive detection of SARS-CoV-2 close to the detection limits of RT-qPCR. Combined RT-qPCR/pyrosequencing does not negatively affect preceding RT-qPCR pipeline in SARS-CoV-2 diagnostics and can be optionally applied in routine to inspect conflicting RT-qPCR results.
RESUMO
Hip range of motion after total hip arthroplasty has been shown to be dependent on prosthetic design and component placement. We hypothesized that bony anatomy would significantly affect range of motion. Computer models of a current generation hip arthroplasty design were virtually implanted in a model of pelvis and femur in various orientations ranging from 35 degrees to 55 degrees cup abduction, 0 degrees to 30 degrees cup anteversion, and 0 degrees to 30 degrees femoral anteversion. Four head sizes ranging from 22.2 to 32 mm and two neck sizes ranging from 10-mm and 12-mm diameter were tested. Range of motion was recorded as maximum flexion-extension, abduction-adduction, and axial rotation of the femur before any contact between prosthetic components or bone was detected. Bony impingement preceded component impingement in about 44% of all conditions tested, ranging from 66% in adduction to 22% in extension. Range of motion increased as head size increased. However, increasing head size also increased the propensity for bony impingement, which tended to reduce the beneficial effect of increased head size on range of motion. Reducing neck diameter had a greater effect on prosthetic impingement (mean, 3.5 degrees increase in range of motion) compared to bone impingement (mean, 1.9 degrees ). This model allowed for a clinically relevant assessment of range of motion after total hip arthroplasty and may also be used with patient-specific geometry [such as that obtained from preoperative computed tomography (CT) scans] for more accurate preoperative planning.
