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Effect of Heat inactivation on Real-Time Reverse Transcription PCR of the SARS-COV-2 Detection
dc.contributor.author | liu, yanxia et al. | |
dc.date.accessioned | 2020-06-16T17:14:20Z | |
dc.date.available | 2020-06-16T17:14:20Z | |
dc.date.issued | 2020-05-28 | |
dc.identifier.uri | https://doi.org/10.1101/2020.05.19.20101469 | en_US |
dc.identifier.uri | https://hdl.handle.net/20.500.12663/1759 | |
dc.description.abstract | Background: Real-time reverse transcription PCR (rRT-PCR) is commonly used to diagnose SARS-CoV-2 infection. Heat inactivation prior to nucleic acid isolation may allow safe testing, while the effects of heat inactivation on SARS-CoV-2 rRT-PCR detection result need to be determined. Methods: 14 positive nasopharyngeal swab specimens were inactivated at 56°C for 30min, 56°C for 60min, 60°C for 30min, 60°C for 75min, and 100°C for 10min, and another 2 positive nasopharyngeal swab specimens were also inactivated at 100°C for 10min, 100°C for 30min, 100°C for 60min, after which the samples were isolated and detected by rRT-PCR. Results: All 14 heat treated samples remained positive. The range of threshold cycle (Ct) values observed when detecting ORF1a/b was 27.228-34.011 in heat-treated samples, while 25.281-34.861 in unheated samples, and the range of threshold cycle (Ct) values observed at the time of detecting N was 25.777-33.351 in heat-treated samples, while 24.1615-35.433 in unheated samples, on basis of which it showed no statistical difference otherwise a good correlation of Ct values between the heat-inactivated samples and the untreated samples. However, the 2 samples inactivated at 100°C 30min, 100°C 60min turned into negative. Conclusions: Heat inactivation at 56°C for 30min, 56°C for 60min, 60°C for 30min, 60°C for 75min, and 100°C for 10min shall not affect the detection results of Real-Time Reverse Transcription PCR of the SARS-COV2. Furthermore, it is recommended to inactive nasopharyngeal swab specimens 10min at 100°C before RNA extraction in consideration of efficiency and reliable results. Key Words: SARS-CoV-2, Heat Inactivation, rRT-PCR, Comparison. | en_US |
dc.language | English | en_US |
dc.subject | COVID-19 | en_US |
dc.subject | Coronavirus | en_US |
dc.subject | Infectious Diseases | en_US |
dc.subject | Betacoronavirus | en_US |
dc.subject | Real-Time Polymerase Chain Reaction | en_US |
dc.subject | Coronavirus Infections | en_US |
dc.title | Effect of Heat inactivation on Real-Time Reverse Transcription PCR of the SARS-COV-2 Detection | en_US |
eihealth.country | Global (WHO/OMS) | en_US |
eihealth.category | Virus: natural history, transmission and diagnostics | en_US |
eihealth.type | Published Article | en_US |
eihealth.maincategory | Slow Spread / Reducir la Dispersión | en_US |
dc.relation.ispartofjournal | medRxiv | en_US |
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