Effect of RNA quality to SARS-CoV-2 RT-qPCR detection from saliva

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http://hdl.handle.net/10138/343821

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Paju , S , Tallgren , M , Kivimäki , A , Lahdentausta , L , Salminen , A , Oksanen , L-M A H , Sanmark , E , Geneid , A , Pussinen , P & Pietiäinen , M 2022 , ' Effect of RNA quality to SARS-CoV-2 RT-qPCR detection from saliva ' , Journal of Medical Microbiology , vol. 71 , no. 4 , 001507 . https://doi.org/10.1099/jmm.0.001507

Title: Effect of RNA quality to SARS-CoV-2 RT-qPCR detection from saliva
Author: Paju, Susanna; Tallgren, Marika; Kivimäki, Anne; Lahdentausta, Laura; Salminen, Aino; Oksanen, Lotta-Maria Adele Helena; Sanmark, Enni; Geneid, Ahmed; Pussinen, Pirkko; Pietiäinen, Milla
Contributor organization: HUS Head and Neck Center
Department of Oral and Maxillofacial Diseases
Clinicum
Department of Pharmacology
Medicum
Department of Ophthalmology and Otorhinolaryngology
Korva-, nenä- ja kurkkutautien klinikka
Faculty Common Matters
Date: 2022
Language: eng
Number of pages: 6
Belongs to series: Journal of Medical Microbiology
ISSN: 0022-2615
DOI: https://doi.org/10.1099/jmm.0.001507
URI: http://hdl.handle.net/10138/343821
Abstract: Saliva is an alternative sample material to nasopharyngeal swab in SARS- CoV- 2 diagnostics. We investigated possible aspects to improve the reliability of SARS- CoV- 2 detection from saliva. Saliva was collected from asymptomatic healthy subjects (n=133) and COVID- 19 patients (n=9). SARS- CoV- 2 detection was performed with quantitative reverse- transcriptase PCR (RT- qPCR) with two viral and one host target serving as an internal control. The use of internal control revealed that in the first RT- qPCR run 25???30 % of assays failed. The failure is associated with poor RNA quality. When the amount of RNA was cut down to half from the original amount, the performance of RT- qPCR was greatly enhanced (95 % of the assays succeeded). The quality of RNA was not affected by the use of different nucleic acid stabilizing buffers. Our study showed that saliva is suitable material for RT- qPCR based SARS- CoV- 2 diagnostics, but the use of internal control is essential to distinguish the true negative samples from failed assays.
Subject: 11832 Microbiology and virology
susanna
paju@helsinki
fi SARS-CoV-2
COVID-19
RT-qPCR
saliva
Peer reviewed: Yes
Rights: cc_by
Usage restriction: openAccess
Self-archived version: publishedVersion


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