Comparison of different methods of measuring angle of progression in prediction of labor outcome

Objective: First, to compare the manual sagittal and para-sagittal and automated para-sagittal methods of measuring the angle of progression (AOP) by transperineal ultrasound during labor, and second, to develop models for the prediction of time-to-delivery and need for cesarean section (CS) for failure to progress (FTP) in a population of patients undergoing induction of labor. Methods: This was a prospective observational study of transperineal ultrasound on a cohort of 512 women with singleton pregnancies undergiong induction of labor. A random selection of 50 stored images was assessed for inter- and intra-observer reliability between methods. In the cases of vaginal delivery univariate linear, multivariate linear and quantile regression were performed to predict time-to-delivery. Univariate and multivariate binomial logistic regression were performed to predict CS for FTP in the first stage of labor. Results: The intra correlation coefficients (ICC) for the manual para-sagittal method for a single observer was 0.97 (CI 0.95-0.98) and for two observers was 0.96 (CI 0.93-0.98) indicating good reliability. The ICC for the sagittal method for a single observer was 0.93 (0.88-0.96) and for two observers was 0.74 (0.58-0.84) indicating moderate reliabilty for a single observer and poor reliability between two observers. Bland-Altman analysis demonstrated narrower limits of agreement for the manual para-saggittal approach than for the sagittal approach for both single and two observers. The automated para-sagittal method failed to capture an image in 19% of cases. The mean difference between sagittal and para-sagittal methods was 110. In pregnancies resulting in vaginal delivery, 54% of the variation in time-to-delivery was explained in a model combining parity, epidural and syntocinon use during labour and the sonographic findings of fetal head position and AOP. In the prediction of CS for FTP in the first stage of labour a model which combined maternal factors with the sonographic measurements of AOP and estimated fetal weight was superior to one utilising maternal factors alone (area under the curve 0.80 vs 0.76). Conclusions: First, the method of measuring AOP with greatest reliability is the manual parasagittal technique and future research should focus on this technique, second, over half of the variation in time to vaginal delivery can be explained by a model that combines maternal factors, pregnancy characteristics and ultrasound findings, and third, the ability of AOP to provide clinically useful prediction CS for FTP in the first stage of labour is limited

Sequential development of several RT-qPCR tests using LNA nucleotides and dual probe technology to differentiate SARS-CoV-2 from influenza A and B

Sensitive and accurate RT-qPCR tests are the primary diagnostic tools to identify SARS-CoV-2-infected patients. While many SARS-CoV-2 RT-qPCR tests are available, there are significant differences in test sensitivity, workflow (e.g. hands-on-time), gene targets and other functionalities that users must consider. Several publicly available protocols shared by reference labs and public health authorities provide useful tools for SARS-CoV-2 diagnosis, but many have shortcomings related to sensitivity and laborious workflows. Here, we describe a series of SARS-CoV-2 RT-qPCR tests that are originally based on the protocol targeting regions of the RNA-dependent RNA polymerase (RdRp) and envelope (E) coding genes developed by the Charite Berlin. We redesigned the primers/probes, utilized locked nucleic acid nucleotides, incorporated dual probe technology and conducted extensive optimizations of reaction conditions to enhance the sensitivity and specificity of these tests. By incorporating an RNase P internal control and developing multiplexed assays for distinguishing SARS-CoV-2 and influenza A and B, we streamlined the workflow to provide quicker results and reduced consumable costs. Some of these tests use modified enzymes enabling the formulation of a room temperature-stable master mix and lyophilized positive control, thus increasing the functionality of the test and eliminating cold chain shipping and storage. Moreover, a rapid, RNA extraction-free version enables high sensitivity detection of SARS-CoV-2 in about an hour using minimally invasive, self-collected gargle samples. These RT-qPCR assays can easily be implemented in any diagnostic laboratory and can provide a powerful tool to detect SARS-CoV-2 and the most common seasonal influenzas during the vaccination phase of the pandemic