The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 remains essential for tracking and containing the rapid spread of the virus. However, due to increased global demand, kits and proprietary reagents for RNA extraction are limited, which markedly reduce SARS-CoV-2 testing capabilities in many countries. Here, we explore the use of conventional acid guanidinium thiocyanate-phenol-chloroform (AGPC)-based RNA purification as an alternative to commercial automated systems for detection of SARS-CoV-2 by RT-qPCR. 87 clinical oropharyngeal or nasopharyngeal swab specimens were extracted by AGPC and compared to the commercial platforms, the Maxwell® RSC 48 instrument for automated RNA extraction and the fully integrated diagnostic system, the Cobas®6800 apparatus. Our results show that RNA extracted using the AGPC method is fully comparable to modern automated systems regarding analytical sensitivity, specificity and accuracy with respect to detection of SARS-CoV-2 as evaluated by RT-qPCR. Moreover, we find that the AGPC method is easily scalable and implemented in conventional laboratories. Taken together, these data identify conventional AGPC-based RNA extraction as a low cost and suitable alternative to automated systems for the detection of SARS-CoV-2, when automated systems, kits and reagents are not readily available.
The outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly reached pandemic levels. Sufficient testing for SARS-CoV-2 has remained essential for tracking and containing the virus. SARS-CoV-2 testing capabilities are still limited in many countries. Here, we explore the use of conventional RNA purification as an alternative to automated systems for detection of SARS-CoV-2 by RT-qPCR. 87 clinical swab specimens were extracted by conventional phenol-chloroform RNA purification and compared to commercial platforms for RNA extraction and the fully integrated Cobas®6800 diagnostic system. Our results show that the conventional RNA extraction is fully comparable to modern automated systems regarding analytical sensitivity and specificity with respect to detection of SARS-CoV-2 as evaluated by RT-qPCR. Moreover, the method is easily scalable and implemented in conventional laboratories as a low cost and suitable alternative to automated systems for the detection of SARS-CoV-2.
This investigation was carried out in an effort to analyze the influence of various degrees of ischaemic heart disease (IHD) on cardiovascular and physical performance. Assessment of the severity of IHD was based on observations obtained routinely during exercise tests (ST segment response and systolic blood pressure response). The study group included 926 subjects with known or suspected IHD, who were referred for an exercise testing; 268 females, mean age 54 years (range 19-89 years), and 658 males, mean age 52 years (range 16-88 years). We found that increasing IHD severity was associated with significant reductions of cardiovascular performance. The mean maximum work-load (MWL) was lower in females than in males, and MWL as well as mean maximum heart rate (MHR) and mean maximum change in systolic blood pressure (M delta SBP) decreased with increasing IHD and age. The present results may be used to assess the cardiovascular response to exercise in patients with IHD so that altered responses due to causes other than IHD may be identified. Furthermore the result may prove useful in the adjustment of rate responsive pacemakers (RRP) in patients with IHD.
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