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Loop-mediated isothermal amplification for the detection of SARS-CoV-2 in saliva.
Janíková, Monika; Hodosy, Július; Boor, Peter; Klempa, Boris; Celec, Peter.
  • Janíková M; Institute of Molecular Biomedicine, Faculty of Medicine, Comenius University, Bratislava, Slovakia.
  • Hodosy J; Institute of Molecular Biomedicine, Faculty of Medicine, Comenius University, Bratislava, Slovakia.
  • Boor P; University Hospital, Bratislava, Slovakia.
  • Klempa B; Institute of Pathology, Department of Nephrology, University Clinic of the RWTH, Aachen, Germany.
  • Celec P; Institute of Virology, Biomedical Research Center, Slovak Academy of Sciences, Bratislava, Slovakia.
Microb Biotechnol ; 14(1): 307-316, 2021 01.
Article in English | MEDLINE | ID: covidwho-1048485
ABSTRACT
In the fight against the recent COVID-19 pandemics, testing is crucial. Nasopharyngeal swabs and real-time RT-PCR are used for the detection of the viral RNA. The collection of saliva is non-invasive, pain-free and does not require trained personnel. An alternative to RT-PCR is loop-mediated isothermal amplification coupled with reverse transcription (RT-LAMP) that is easy to perform, quick and does not require a thermal cycler. The aim of this study was to test whether SARS-CoV-2 RNA can be detected directly in saliva using RT-LAMP. We have tested 16 primer mixes from the available literature in three rounds of sensitivity assays. The selected RT-LAMP primer mix has a limit of detection of 6 copies of viral RNA per reaction in comparison with RT-PCR with 1 copy per reaction. Whole saliva, as well as saliva collected using Salivette collection tubes, interfered with the RT-LAMP analysis. Neither Chelex-100 nor protease treatment of saliva prevented the inhibitory effect of saliva. With the addition of the ribonuclease inhibitor, the sensitivity of the RT-LAMP assay was 12 copies per reaction of RNA in Salivette® saliva samples and 6 copies per reaction of RNA in whole saliva samples. This study shows that it is possible to combine the use of saliva and RT-LAMP for SARS-CoV-2 RNA detection without RNA extraction which was confirmed on a small set of correctly diagnosed clinical samples. Further studies should prove whether this protocol is suitable for point of care testing in the clinical setting.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Saliva / Nucleic Acid Amplification Techniques / Molecular Diagnostic Techniques / COVID-19 Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Microb Biotechnol Year: 2021 Document Type: Article Affiliation country: 1751-7915.13737

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Saliva / Nucleic Acid Amplification Techniques / Molecular Diagnostic Techniques / COVID-19 Testing / SARS-CoV-2 / COVID-19 Type of study: Diagnostic study / Prognostic study Limits: Humans Language: English Journal: Microb Biotechnol Year: 2021 Document Type: Article Affiliation country: 1751-7915.13737