Direct-RT-qPCR detection of SARS-CoV-2 without RNA extraction as part of a COVID-19 testing strategy: from sample to result in one hour

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61989592%3A15110%2F20%3A73601301" target="_blank" >RIV/61989592:15110/20:73601301 - isvavai.cz</a>

Alternative codes found

RIV/00098892:_____/20:N0000142

Result on the web

<a href="https://www.mdpi.com/2075-4418/10/8/605/htm" target="_blank" >https://www.mdpi.com/2075-4418/10/8/605/htm</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.3390/diagnostics10080605" target="_blank" >10.3390/diagnostics10080605</a>

Result language

angličtina

Original language name

Direct-RT-qPCR detection of SARS-CoV-2 without RNA extraction as part of a COVID-19 testing strategy: from sample to result in one hour

Original language description

Due to the lack of protective immunity in the general population and the absence of effective antivirals and vaccines, the COVID-19 pandemic continues in some countries, with local epicentres emerging in others. Due to the great demand for effective COVID-19 testing programmes to control the spread of the disease, we have suggested such a testing programme that includes a rapid RT-qPCR approach without RNA extraction. The Direct-One-Step-RT-qPCR (DIOS-RT-qPCR) assay detects SARS-CoV-2 in less than one hour while maintaining the high sensitivity and specificity required of diagnostic tools. This optimised protocol allows for the direct use of swab transfer media (14 l) without the need for RNA extraction, achieving comparable sensitivity to the standard method that requires the time-consuming and costly step of RNA isolation. The limit of detection for DIOS-RT-qPCR was lower than seven copies/reaction, which translates to 550 virus copies/ml of swab. The speed, ease of use and low price of this assay make it suitable for high-throughput screening programmes. The use of fast enzymes allows RT-qPCR to be performed under standard laboratory conditions within one hour, making it a potential point-of-care solution on high-speed cycling instruments. This protocol also implements the heat inactivation of SARS-CoV-2 (75°C for 10 min), which renders samples non-infectious, enabling testing in BSL-2 facilities. Moreover, we discuss the critical steps involved in developing tests for the rapid detection of COVID-19. Implementing rapid, easy, cost-effective methods can help control the worldwide spread of the COVID-19 infection.

Czech name

—

Czech description

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Type

J_SC - Article in a specialist periodical, which is included in the SCOPUS database

CEP classification

—

OECD FORD branch

30102 - Immunology

Project

<a href="/en/project/LM2010004" target="_blank" >LM2010004: BBMRI_CZ in the building of the Czech part of a large distributed research infrastructure of pan-European importance: the creation and operation of the network of banks of biological material for biomedical research</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>S - Specificky vyzkum na vysokych skolach<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2020

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Diagnostics

ISSN

2075-4418

e-ISSN

—

Volume of the periodical

10

Issue of the periodical within the volume

8

Country of publishing house

CH - SWITZERLAND

Number of pages

10

Pages from-to

1-10

UT code for WoS article

000564746000001

EID of the result in the Scopus database

2-s2.0-85090354620