The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) outbreak urgently necessitates sensitive and convenient COVID-19 diagnostics for the containment and timely treatment of patients. We aimed to develop and validate a novel reverse transcription–loop-mediated isothermal amplification (RT-LAMP) assay to detect SARS-CoV-2. Patients with suspected COVID-19 and close contacts were recruited from two hospitals between 26 January and 8 April 2020. Respiratory samples were collected and tested using RT-LAMP, and the results were compared with those obtained by reverse transcription-quantitative PCR (RT-qPCR). Samples yielding inconsistent results between these two methods were subjected to next-generation sequencing for confirmation. RT-LAMP was also applied to an asymptomatic COVID-19 carrier and patients with other respiratory viral infections. Samples were collected from a cohort of 129 cases (329 nasopharyngeal swabs) and an independent cohort of 76 patients (152 nasopharyngeal swabs and sputum samples). The RT-LAMP assay was validated to be accurate (overall sensitivity and specificity of 88.89% and 99.00%, respectively) and diagnostically useful (positive and negative likelihood ratios of 88.89 and 0.11, respectively). RT-LAMP showed increased sensitivity (88.89% versus 81.48%) and high consistency (kappa, 0.92) compared to those of RT-qPCR for SARS-CoV-2 screening while requiring only constant-temperature heating and visual inspection. The time required for RT-LAMP was less than 1 h from sample preparation to the result. In addition, RT-LAMP was feasible for use with asymptomatic patients and did not cross-react with other respiratory pathogens. The developed RT-LAMP assay offers rapid, sensitive, and straightforward detection of SARS-CoV-2 infection and may aid the expansion of COVID-19 testing in the public domain and hospitals. IMPORTANCE We developed a visual and rapid reverse transcription–loop-mediated isothermal amplification (RT-LAMP) assay targeting the S gene for SARS-CoV-2 infection. The strength of our study was that we validated the RT-LAMP assay using 481 clinical respiratory samples from two prospective cohorts of suspected COVID-19 patients and on the serial samples from an asymptomatic carrier. The developed RT-LAMP approach showed an increased sensitivity (88.89%) and high consistency (kappa, 0.92) compared with those of reverse transcription-quantitative PCR (RT-qPCR) for SARS-CoV-2 screening while requiring only constant-temperature heating and visual inspection, facilitating SARS-CoV-2 screening in well-equipped labs as well as in the field. The time required for RT-LAMP was less than 1 h from sample preparation to the result (more than 2 h for RT-qPCR). This study showed that the RT-LAMP assay was a simple, rapid, and sensitive approach for SARS-CoV-2 infection and can facilitate COVID-19 diagnosis, especially in resource-poor settings.
For a Legendrian (2, n) torus knot or link with maximal Thurston-Bennequin number, Ekholm, Honda, and Kálmán [EHK] constructed C n exact Lagrangian fillings, where C n is the n-th Catalan number. We show that these exact Lagrangian fillings are pairwise non-isotopic through exact Lagrangian isotopy. To do that, we compute the augmentations induced by the exact Lagrangian fillings L to Z 2 [H 1 (L)] and distinguish the resulting augmentations.
For 1-dimensional Legendrian submanifolds of 1-jet spaces, we extend the functorality of the Legendrian contact homology DGalgebra (DGA) from embedded exact Lagrangian cobordisms, as in [16], to a class of immersed exact Lagrangian cobordisms by considering their Legendrian lifts as conical Legendrian cobordisms. To a conical Legendrian cobordism Σ from Λ − to Λ + , we associate an immersed DGA map, which is a diagramwhere f is a DGA map and i is an inclusion map. This construction gives a functor between suitably defined categories of Legendrians with immersed Lagrangian cobordisms and DGAs with immersed DGA maps. In an algebraic preliminary, we consider an analog of the mapping cylinder construction in the setting of DG-algebras and establish several of its properties. As an application we give examples of augmentations of Legendrian twist knots that can be induced by an immersed filling with a single double point but cannot be induced by any orientable embedded filling. Y. Pan and D. Rutherford 6 The immersed LCH functor I: Construction of immersed maps 687 7 The immersed LCH functor II: Proof of statements 698 8 Augmentations from immersed Lagrangian fillings 706 9 The SFT perspective 714 References 719Functorial LCH for immersed Lagrangian cobordisms 643 Finally in Section 9, we translate the story to Symp(J 1 M ) and show that it matches with the SFT framework. Acknowledgements
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