Development and implementation of a RT‐qPCR extraction‐free protocol for the detection of SARS‐CoV‐2 and impact on the turn‐around‐time
Abstract The occurrence of the COVID‐19 second‐wave outbreak in Europe has pushed laboratories performing molecular SARS‐CoV‐2 tests to increase their throughput and decrease the result rendering time. In this evaluation, we tested for the first time a new, extraction‐free, protocol with the Allplex SARS‐CoV‐2 Assay RT‐qPCR kit on a Nimbus platform. Ninety‐one samples, of which 71 previously tested positive with RT‐qPCR with extraction were immediately analyzed without extraction, using only a dilution and thermal shock protocol. The positive and negative percentage agreements were respectively 97.2% (95% confidence interval [CI]: 0.90–0.99) and 95.0% (95% CI: 0.76–0.99). The two false negatives observed were very weakly positive with the comparison method. Moderate variations in Ct of the targeted genes were observed (median ± 95% CI): E gene, +2.49 ± 0.44; N gene, +0.98 ± 0.54; RdRP/S genes, +2.64 ± 0.48. On the other hand, the number of tests performed within 24 h raised from 86.4% to 97.8%, the turn‐around time decreased from 19:18 to 09:03 ( p < .0001), and the number of tests that can be performed per day doubled since this technique was introduced routinely in our laboratory..
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:93 |
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| Enthalten in: |
Journal of Medical Virology - 93(2021), 4, Seite 2538-2542 |
| Beteiligte Personen: |
Blairon, Laurent [VerfasserIn] |
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| BKL: |
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| Anmerkungen: |
© 2021 Wiley Periodicals LLC |
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| Umfang: |
5 |
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| doi: |
10.1002/jmv.26782 |
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| funding: |
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| 520 | |a Abstract The occurrence of the COVID‐19 second‐wave outbreak in Europe has pushed laboratories performing molecular SARS‐CoV‐2 tests to increase their throughput and decrease the result rendering time. In this evaluation, we tested for the first time a new, extraction‐free, protocol with the Allplex SARS‐CoV‐2 Assay RT‐qPCR kit on a Nimbus platform. Ninety‐one samples, of which 71 previously tested positive with RT‐qPCR with extraction were immediately analyzed without extraction, using only a dilution and thermal shock protocol. The positive and negative percentage agreements were respectively 97.2% (95% confidence interval [CI]: 0.90–0.99) and 95.0% (95% CI: 0.76–0.99). The two false negatives observed were very weakly positive with the comparison method. Moderate variations in Ct of the targeted genes were observed (median ± 95% CI): E gene, +2.49 ± 0.44; N gene, +0.98 ± 0.54; RdRP/S genes, +2.64 ± 0.48. On the other hand, the number of tests performed within 24 h raised from 86.4% to 97.8%, the turn‐around time decreased from 19:18 to 09:03 ( p < .0001), and the number of tests that can be performed per day doubled since this technique was introduced routinely in our laboratory. | ||
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