An in-well direct lysis method for rapid detection of SARS-CoV-2 by real time RT-PCR in eSwab specimens.
J Virol Methods
; 289: 114062, 2021 03.
Article
in English
| MEDLINE | ID: covidwho-1019346
ABSTRACT
BACKGROUND:
Diagnostic real time reverse transcription PCR (rRT-PCR) is usually done using nucleic acid (NA) purified from the sample. In the SARS-CoV-2 pandemic reagents and utensils for NA purification has been in short supply. This has generated interest in methods that eliminate the need for NA purification.OBJECTIVES:
To investigate if addition of detergent to rRT-PCR master mix (MM) enabled in-well direct lysis and detection of SARS-CoV-2 in clinical eSwab specimens. STUDYDESIGN:
IGEPAL-CA-630 (IGEPAL) was added to SARS-CoV-2 MM to 0.3 % final concentration and crude sample was added directly to the PCR well containing MM. Cycle of positivity (Cp) and categorical agreement was compared in samples tested in standard rRT-PCR after NA purification and in in-well lysis, direct rRT-PCR.RESULTS:
In-well lysis direct rRT-PCR detected SARS-CoV-2 in 27/30 previously SARS-CoV-2+ samples with an average bias of 3.26 cycles (95 %CI 0.08-6.43 cycles). All 30 previously test negative samples remained negative when tested in in-well lysis, direct PCR.CONCLUSIONS:
Supplementation of detergent to MM was shown to be useful for the detection of SARS CoV-2 in eSwab specimens (COPAN) by direct rRT-PCR without prior NA purification.Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Specimen Handling
/
RNA, Viral
/
COVID-19 Nucleic Acid Testing
/
SARS-CoV-2
/
COVID-19
Type of study:
Diagnostic study
/
Prognostic study
Limits:
Humans
Language:
English
Journal:
J Virol Methods
Year:
2021
Document Type:
Article
Affiliation country:
J.jviromet.2021.114062
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