Analytical and clinical evaluation of a heat shock SARS-CoV-2 detection method without RNA extraction for N and E genes RT-qPCR
Background: The COVID-19 pandemic has caused significant supply shortages worldwide for SARS-CoV-2 molecular diagnosis, like RNA extraction kits. Objective: The aim of our study was to evaluate the clinical performance and analytical sensitivity of a simple SARS-CoV-2 diagnosis protocol based on heat shock without RNA extraction using both ''CDC'' (N gene) and ''Charite'' (E gene) RT-qPCR protocols. Results: 1,036 nasopharyngeal samples, 543 of them SARS-CoV-2 positive, were analyzed. The heat shock method correctly identified 68.8% (232/337) and 89.4% (202/226) of SARS-CoV-2 positive samples for N gene and E gene, respectively. Analytical sensitivity was assessed for heat shock method using the CDC RT-qPCR protocol, obtaining sensitivity values of 98.6%, 93.3% and 84.8% for limit of detection of 100.000, 50.000 and 20.000 viral RNA copies/mL of sample. Conclusions: Our findings show that a simple heat shock SARS-CoV-2 RT-qPCR diagnosis method without RNA extraction is a reliable alternative for potentially infectious SARS-CoV-2 positive patients at the time of testing. This affordable protocol can help overcome the cost and supply shortages for SARS-CoV-2 diagnosis, especially in developing countries. In Ecuador, it has been used already by laboratories in the public health system for more than 100.000 specimens..
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2021 |
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| Erschienen: |
2021 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:109 |
|---|---|
| Enthalten in: |
International Journal of Infectious Diseases - 109(2021), Seite 315-320 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Alfredo Bruno [VerfasserIn] |
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| Links: |
doi.org [kostenfrei] |
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| Themen: |
COVID-19 |
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| doi: |
10.1016/j.ijid.2021.06.038 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
DOAJ049837532 |
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| 520 | |a Background: The COVID-19 pandemic has caused significant supply shortages worldwide for SARS-CoV-2 molecular diagnosis, like RNA extraction kits. Objective: The aim of our study was to evaluate the clinical performance and analytical sensitivity of a simple SARS-CoV-2 diagnosis protocol based on heat shock without RNA extraction using both ''CDC'' (N gene) and ''Charite'' (E gene) RT-qPCR protocols. Results: 1,036 nasopharyngeal samples, 543 of them SARS-CoV-2 positive, were analyzed. The heat shock method correctly identified 68.8% (232/337) and 89.4% (202/226) of SARS-CoV-2 positive samples for N gene and E gene, respectively. Analytical sensitivity was assessed for heat shock method using the CDC RT-qPCR protocol, obtaining sensitivity values of 98.6%, 93.3% and 84.8% for limit of detection of 100.000, 50.000 and 20.000 viral RNA copies/mL of sample. Conclusions: Our findings show that a simple heat shock SARS-CoV-2 RT-qPCR diagnosis method without RNA extraction is a reliable alternative for potentially infectious SARS-CoV-2 positive patients at the time of testing. This affordable protocol can help overcome the cost and supply shortages for SARS-CoV-2 diagnosis, especially in developing countries. In Ecuador, it has been used already by laboratories in the public health system for more than 100.000 specimens. | ||
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| 653 | 0 | |a Infectious and parasitic diseases | |
| 700 | 0 | |a Domenica de Mora |e verfasserin |4 aut | |
| 700 | 0 | |a Byron Freire-Paspuel |e verfasserin |4 aut | |
| 700 | 0 | |a Angel S. Rodriguez |e verfasserin |4 aut | |
| 700 | 0 | |a Maria Belen Paredes-Espinosa |e verfasserin |4 aut | |
| 700 | 0 | |a Maritza Olmedo |e verfasserin |4 aut | |
| 700 | 0 | |a Martha Sanchez |e verfasserin |4 aut | |
| 700 | 0 | |a Jennifer Romero |e verfasserin |4 aut | |
| 700 | 0 | |a Michelle Paez |e verfasserin |4 aut | |
| 700 | 0 | |a Manuel Gonzalez |e verfasserin |4 aut | |
| 700 | 0 | |a Alberto Orlando |e verfasserin |4 aut | |
| 700 | 0 | |a Miguel Angel Garcia-Bereguiain |e verfasserin |4 aut | |
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