tetano
Editor, Senior Moderator
Funct Integr Genomics
. 2022 Sep 12.
doi: 10.1007/s10142-022-00900-5. Online ahead of print.
Colorimetric and fluorometric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for diagnosis of SARS-CoV-2
Galyah Alhamid[SUP] 1 2 [/SUP], Huseyin Tombuloglu[SUP] 3 [/SUP], Dalal Motabagani[SUP] 4 [/SUP], Dana Motabagani[SUP] 4 [/SUP], Ali A Rabaan[SUP] 5 6 7 [/SUP], Kubra Unver[SUP] 8 [/SUP], Gabriel Dorado[SUP] 9 [/SUP], Ebtesam Al-Suhaimi[SUP] 10 [/SUP], Turgay Unver[SUP] 11 [/SUP]
Affiliations
Abstract
The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused millions of infections and deaths worldwide since it infected humans almost 3 years ago. Improvements of current assays and the development of new rapid tests or to diagnose SARS-CoV-2 are urgent. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a rapid and propitious assay, allowing to detect both colorimetric and/or fluorometric nucleic acid amplifications. This study describes the analytical and clinical evaluation of RT-LAMP assay for detection of SARS-CoV-2, by designing LAMP primers targeting N (nucleocapsid phosphoprotein), RdRp (polyprotein), S (surface glycoprotein), and E (envelope protein) genes. The assay's performance was compared with the gold standard RT-PCR, yielding 94.6% sensitivity and 92.9% specificity. Among the tested primer sets, the ones for S and N genes had the highest analytical sensitivity, showing results in about 20 min. The colorimetric and fluorometric comparisons revealed that the latter is faster than the former. The limit of detection (LoD) of RT-LAMP reaction in both assays is 50 copies/µl of the reaction mixture. However, the simple eye-observation advantage of the colorimetric assay (with a color change from yellow to red) serves a promising on-site point-of-care testing method anywhere, including, for instance, laboratory and in-house applications.
Keywords: COVID-19; Colorimetric; Coronavirus; Diagnosis; Fluorometric; RT-LAMP; SARS-CoV-2.
. 2022 Sep 12.
doi: 10.1007/s10142-022-00900-5. Online ahead of print.
Colorimetric and fluorometric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for diagnosis of SARS-CoV-2
Galyah Alhamid[SUP] 1 2 [/SUP], Huseyin Tombuloglu[SUP] 3 [/SUP], Dalal Motabagani[SUP] 4 [/SUP], Dana Motabagani[SUP] 4 [/SUP], Ali A Rabaan[SUP] 5 6 7 [/SUP], Kubra Unver[SUP] 8 [/SUP], Gabriel Dorado[SUP] 9 [/SUP], Ebtesam Al-Suhaimi[SUP] 10 [/SUP], Turgay Unver[SUP] 11 [/SUP]
Affiliations
- PMID: 36089609
- DOI: 10.1007/s10142-022-00900-5
Abstract
The coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has caused millions of infections and deaths worldwide since it infected humans almost 3 years ago. Improvements of current assays and the development of new rapid tests or to diagnose SARS-CoV-2 are urgent. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) is a rapid and propitious assay, allowing to detect both colorimetric and/or fluorometric nucleic acid amplifications. This study describes the analytical and clinical evaluation of RT-LAMP assay for detection of SARS-CoV-2, by designing LAMP primers targeting N (nucleocapsid phosphoprotein), RdRp (polyprotein), S (surface glycoprotein), and E (envelope protein) genes. The assay's performance was compared with the gold standard RT-PCR, yielding 94.6% sensitivity and 92.9% specificity. Among the tested primer sets, the ones for S and N genes had the highest analytical sensitivity, showing results in about 20 min. The colorimetric and fluorometric comparisons revealed that the latter is faster than the former. The limit of detection (LoD) of RT-LAMP reaction in both assays is 50 copies/µl of the reaction mixture. However, the simple eye-observation advantage of the colorimetric assay (with a color change from yellow to red) serves a promising on-site point-of-care testing method anywhere, including, for instance, laboratory and in-house applications.
Keywords: COVID-19; Colorimetric; Coronavirus; Diagnosis; Fluorometric; RT-LAMP; SARS-CoV-2.