METHODS: SARS-CoV-2 antigens were immobilized on nitrocellulose membrane to capture human IgG, which was then detected with anti-human IgG conjugated gold nanoparticle (hIgG-AuNP). A total of 181 samples were analyzed in-house. Within which 35 were further evaluated in US FDA-approved CLIA Elecsys SARS-CoV-2 assay. The positive panel consisted of RT-qPCR positive samples from patients with both <14 days and >14 days from the onset of clinical symptoms. The negative panel contained samples collected from the pre-pandemic era dengue patients and healthy donors during the pandemic. Moreover, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of FT-DBA were evaluated against RT-qPCR positive sera. However, the overall efficacies were assessed with sera that seroconverted against either nucleocapsid (NCP) or receptor-binding domain (RBD).
RESULTS: In-house ELISA selected a total of 81 true seropositive and 100 seronegative samples. The sensitivity of samples with <14 days using FT-DBA was 94.7%, increasing to 100% for samples >14 days. The overall detection sensitivity and specificity were 98.8% and 98%, respectively, whereas the overall PPV and NPV were 99.6% and 99%. Moreover, comparative analysis between in-house ELISA assays and FT-DBA revealed clinical agreement of Cohen's Kappa value of 0.944. The FT-DBA showed sensitivity and specificity of 100% when compared with commercial CLIA kits.
CONCLUSION: The assay can confirm past SARS-CoV-2 infection with high accuracy within 2 minutes compared to commercial CLIA or in-house ELISA. It can help track SARS-CoV-2 disease progression, population screening, and vaccination response. The ease of use of the assay without requiring any instruments while being semi-quantitative provides the avenue of its implementation in remote areas around the globe, where conventional serodiagnosis is not feasible.
Materials and Methods: Sixty freshly extracted maxillary incisors teeth collected in saline. Access cavity prepared and canals were made free of bacterial and pulp. The teeth were mounted on the bacteria collecting apparatus. Root canals were contaminated with the Fusobacterium Nucleatum (ATCC25586) and dried at 37°C for 24 h. In Group 1 (Control group): No instrumentation was done and biomechanical preparation done in all other groups with Group 2: Hand K-files, Group 3: Protaper gold, Group 4: K3XF, Group 5: Edge taper platinum, and Group 6: Hyflex CM rotary file systems. Then, the extrude was collected, and it is incubated in Mueller-Hinton agar for 24 h and the number of colony forming units were counted and statistical comparison was done using Kruskal-Wallis test and Mann-Whitney U test.
Results: Hand K-files extruded more bacteria when compared to other four rotary systems, K3XF file system extruded least number of bacteria.
Conclusion: All instrumentation techniques extruded intracanal bacteria apically. However, engine-driven nickel-titanium instruments extruded less bacteria than the manual technique. The K3XF rotary file system comparatively extruded less bacteria than other rotary file systems.