Extraction Bottleneck in the Diagnosis of SARS-CoV-2: Evaluation of an Alternative Protocol Derived from Veterinary Use
Abstract
:1. Introduction
2. Materials and Methods
2.1. Study Design
2.2. Total RNA Extraction
2.3. RT-qPCR
2.4. Data Analysis
3. Results
3.1. Clinical Performance
3.2. Cycle Threshold Analysis
3.3. Limit of Detection
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Testing Method | Reference Method | |
---|---|---|
Positive samples | 59 | 60 |
3/3 genes | 56 | 57 |
2/3 genes | 2 | 2 |
1/3 genes | 1 | 1 |
Negative samples | 13 (+1 *) | 13 |
Total | 73 | 73 |
Overall Sensitivity | 98.33% | |
Overall Specificity | 100% | |
Cohen’s kappa coefficient | 0.955 (excellent) |
Concentration (In TCID50/mL) | Reference Method | Testing Method | ||||
---|---|---|---|---|---|---|
Rep 1 | Rep 2 | Rep 3 | Rep 1 | Rep 2 | Rep 3 | |
1000 | + | + | + | + | + | + |
100 | + | + | + | + | + | + |
10 | + | + | + | + | + | + |
1 | + | + | + | + | + * | + |
0.1 | + | + | + | + | + | + |
0.01 | + | + | + | + | + | + |
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Bottino, P.; Zanotto, E.; Sidoti, F.; Pastrone, L.; Piva, R.; Mereu, E.; Costa, C.; Cavallo, R. Extraction Bottleneck in the Diagnosis of SARS-CoV-2: Evaluation of an Alternative Protocol Derived from Veterinary Use. Microorganisms 2023, 11, 535. https://doi.org/10.3390/microorganisms11020535
Bottino P, Zanotto E, Sidoti F, Pastrone L, Piva R, Mereu E, Costa C, Cavallo R. Extraction Bottleneck in the Diagnosis of SARS-CoV-2: Evaluation of an Alternative Protocol Derived from Veterinary Use. Microorganisms. 2023; 11(2):535. https://doi.org/10.3390/microorganisms11020535
Chicago/Turabian StyleBottino, Paolo, Elisa Zanotto, Francesca Sidoti, Lisa Pastrone, Roberto Piva, Elisabetta Mereu, Cristina Costa, and Rossana Cavallo. 2023. "Extraction Bottleneck in the Diagnosis of SARS-CoV-2: Evaluation of an Alternative Protocol Derived from Veterinary Use" Microorganisms 11, no. 2: 535. https://doi.org/10.3390/microorganisms11020535
APA StyleBottino, P., Zanotto, E., Sidoti, F., Pastrone, L., Piva, R., Mereu, E., Costa, C., & Cavallo, R. (2023). Extraction Bottleneck in the Diagnosis of SARS-CoV-2: Evaluation of an Alternative Protocol Derived from Veterinary Use. Microorganisms, 11(2), 535. https://doi.org/10.3390/microorganisms11020535