Detection of SARS-CoV-2 RNA in nasopharyngeal swabs from COVID-19 patients and asymptomatic cases of infection by real-time and digital PCR.
Обнаружение РНК SARS-CoV-2 в носоглоточных мазках больных COVID-19 и бессимптомных носителей методами цифровой ПЦР и ПЦР в реальном времени.
COVID-19
SARS-CoV-2
digital PCR
real time PCR
Journal
Klinicheskaia laboratornaia diagnostika
ISSN: 0869-2084
Titre abrégé: Klin Lab Diagn
Pays: Russia (Federation)
ID NLM: 9432021
Informations de publication
Date de publication:
29 Dec 2020
29 Dec 2020
Historique:
entrez:
29
12
2020
pubmed:
30
12
2020
medline:
1
1
2021
Statut:
ppublish
Résumé
In this work we tested two reagent kits developed by us for detecting SARS-CoV-2 RNA using a fragment of the ORF1ab gene in digital PCR and real-time PCR formats. Data were obtained on the detection of SARS-CoV-2 virus RNA in nasopharyngeal swabs of patients with COVID-19 and asymptomatic carriers. The developed reagent kits provided 100% sensitivity and a detection limit of 103 GE / ml for qPCR, and at least 200 copies / ml of viral RNA when performing digital PCR. These methods were tested using a panel of 1,328 samples collected from patients with suspected COVID-19 at the beginning of 2020 in the Russian Federation. It has been shown that dPCR is more sensitive and can be used to analyze samples with low viral load, including those from patients without clinical symptoms. dPCR significantly improves the accuracy of laboratory research and significantly reduces the number of false negative results in the diagnosis of SARS-CoV-2. Determination of the concentration of SARS-CoV-2 RNA in patients with different clinical course of the disease showed that the concentration of viral RNA can sharply decrease in the first days of the disease. A low concentration of viral RNA in samples from patients is also characteristic of asymptomatic disease. Digital PCR provides a higher detection rate for asymptomatic cases, which is approximately 75% of those infected, as opposed to 45% for real-time PCR. The results obtained on the use of the digital PCR method for detecting SARS-CoV-2 RNA showed that this method is especially suitable for detecting RNA in case of its low concentration in contacts, as well as for monitoring changes in viral load in convalescent patients.
Identifiants
pubmed: 33373511
doi: 10.18821/0869-2084-2020-65-12-785-792
doi:
Substances chimiques
RNA, Viral
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
785-792Déclaration de conflit d'intérêts
The authors declare no conflict of interest.
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