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AuthorBenslimane, Fatiha M.
AuthorAl-Jamal, Ola
AuthorBoughattas, Sonia|Al Thani, Asmaa A
AuthorYassine, Hadi M.
Available date2020-10-22T06:07:18Z
Publication Date2020
Publication NameQatar University Annual Research an Exhibition 2020 (quarfe)
URIhttps://doi.org/10.29117/quarfe.2020.0288
URIhttp://hdl.handle.net/10576/16512
AbstractBackground: First described 20 years ago by Notomi et al., the loop-mediated isothermal amplification (LAMP) assay is robust, rapid and straightforward, yet retains high sensitivity and specificity. These features have seen the LAMP assay and the inclusion of a reverse transcriptase (RT-LAMP) implemented for a broad range of molecular diagnostic applications extending from infectious diseases, including detection of the original SARS-CoV-1 virus. The advantages of RT-LAMP include using different reagents than RT-qPCR, the potential for direct processing of samples without the need for prior RNA extraction and an extremely rapid turn-around time. Several groups have now described different RT-LAMP assays for detection of SARS-CoV-2 RNA. Therefore, the aim of this study is to assess the feasibility, sensitivity and effectiveness of LAMP technique in detecting SARS-CoV-2 in different type of samples. Method: New England Biolabs (NEB) LAMP master mixes were used. Six set of primers specific to SARS-CoV-2 were obtained from IDT. The reaction mix consisting of LAMP master mix, primer working solution and a sample was incubated at 65?C and results were collected after 30 mins. Results: In just 30 min we were able to detect the virus without any prior sample processing. Our primers were able to detect up to 100 copies of the viruses, which is comparable to the RT-PCR that we currently use in our lab. The primers were tested against all other coronavirus and they have shown 100% specificity to the novel SARS-CoV-2 virus. Both the florescent and calorimetric master mixes were able to detect the virus in all tested samples: clinical, animal and environmental. Conclusion: LAMP is a fast reliable technique that could be used as a quick screening method for the detection of SARS-CoV-2 in different settings and using different collection medium.
Languageen
PublisherQatar University Press
SubjectLAMP
COVID19
SARS-CoV-2
Detection
RT_PCR
TitleEvaluation of reverse transcription-loop-mediated isothermal amplification (RT-LAMP) for detecting SARS-CoV-2 in clinical, environmental and animal samples.
TypePoster
dc.accessType Open Access


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