tetano
Editor, Senior Moderator
Analyst
. 2020 Nov 9.
doi: 10.1039/d0an01775b. Online ahead of print.
Colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a visual diagnostic platform for the detection of the emerging coronavirus SARS-CoV-2
Kawin Nawattanapaiboon[SUP] 1 [/SUP], Ekawat Pasomsub[SUP] 2 [/SUP], Photchanathorn Prombun[SUP] 3 [/SUP], Akanit Wongbunmak[SUP] 3 [/SUP], Akarawit Jenjitwanich[SUP] 3 [/SUP], Pantanat Mahasupachai[SUP] 3 [/SUP], Purichaya Vetcho[SUP] 3 [/SUP], Cholticha Chayrach[SUP] 3 [/SUP], Natthapon Manatjaroenlap[SUP] 3 [/SUP], Chonchanok Samphaongern[SUP] 3 [/SUP], Treewat Watthanachockchai[SUP] 2 [/SUP], Phonthanat Leedorkmai[SUP] 2 [/SUP], Suwimon Manopwisedjaroen[SUP] 4 [/SUP], Radeekorn Akkarawongsapat[SUP] 4 [/SUP], Arunee Thitithanyanont[SUP] 4 [/SUP], Matthew Phanchana[SUP] 5 [/SUP], Watanalai Panbangred[SUP] 6 [/SUP], Somchai Chauvatcharin[SUP] 7 [/SUP], Toemsak Srikhirin[SUP] 8 [/SUP]
Affiliations
Abstract
COVID-19, caused by the infection of SARS-CoV-2, has emerged as a rapidly spreading infection. The disease has now reached the level of a global pandemic and as a result a more rapid and simple detection method is imperative to curb the spread of the virus. We aimed to develop a visual diagnostic platform for SARS-CoV-2 based on colorimetric RT-LAMP with levels of sensitivity and specificity comparable to that of commercial qRT-PCR assays. In this work, the primers were designed to target a conserved region of the RNA-dependent RNA polymerase gene (RdRp). The assay was characterized for its sensitivity and specificity, and validated with clinical specimens collected in Thailand. The developed colorimetric RT-LAMP assay could amplify the target gene and enabled visual interpretation in 60 min at 65 ?C. No cross-reactivity with six other common human respiratory viruses (influenza A virus subtypes H1 and H3, influenza B virus, respiratory syncytial virus types A and B, and human metapneumovirus) and five other human coronaviruses (MERS-CoV, HKU-1, OC43, 229E and NL63) was observed. The limit of detection was 25 copies per reaction when evaluated with contrived specimens. However, the detection rate at this concentration fell to 95.8% when the incubation time was reduced from 60 to 30 min. The diagnostic performance of the developed RT-LAMP assay was evaluated in 2120 clinical specimens and compared with the commercial qRT-PCR. The results revealed high sensitivity and specificity of 95.74% and 99.95%, respectively. The overall accuracy of the RT-LAMP assay was determined to be 99.86%. In summary, our results indicate that the developed colorimetric RT-LAMP provides a simple, sensitive and reliable approach for the detection of SARS-CoV-2 in clinical samples, implying its beneficial use as a diagnostic platform for COVID-19 screening.
. 2020 Nov 9.
doi: 10.1039/d0an01775b. Online ahead of print.
Colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a visual diagnostic platform for the detection of the emerging coronavirus SARS-CoV-2
Kawin Nawattanapaiboon[SUP] 1 [/SUP], Ekawat Pasomsub[SUP] 2 [/SUP], Photchanathorn Prombun[SUP] 3 [/SUP], Akanit Wongbunmak[SUP] 3 [/SUP], Akarawit Jenjitwanich[SUP] 3 [/SUP], Pantanat Mahasupachai[SUP] 3 [/SUP], Purichaya Vetcho[SUP] 3 [/SUP], Cholticha Chayrach[SUP] 3 [/SUP], Natthapon Manatjaroenlap[SUP] 3 [/SUP], Chonchanok Samphaongern[SUP] 3 [/SUP], Treewat Watthanachockchai[SUP] 2 [/SUP], Phonthanat Leedorkmai[SUP] 2 [/SUP], Suwimon Manopwisedjaroen[SUP] 4 [/SUP], Radeekorn Akkarawongsapat[SUP] 4 [/SUP], Arunee Thitithanyanont[SUP] 4 [/SUP], Matthew Phanchana[SUP] 5 [/SUP], Watanalai Panbangred[SUP] 6 [/SUP], Somchai Chauvatcharin[SUP] 7 [/SUP], Toemsak Srikhirin[SUP] 8 [/SUP]
Affiliations
- PMID: 33165486
- DOI: 10.1039/d0an01775b
Abstract
COVID-19, caused by the infection of SARS-CoV-2, has emerged as a rapidly spreading infection. The disease has now reached the level of a global pandemic and as a result a more rapid and simple detection method is imperative to curb the spread of the virus. We aimed to develop a visual diagnostic platform for SARS-CoV-2 based on colorimetric RT-LAMP with levels of sensitivity and specificity comparable to that of commercial qRT-PCR assays. In this work, the primers were designed to target a conserved region of the RNA-dependent RNA polymerase gene (RdRp). The assay was characterized for its sensitivity and specificity, and validated with clinical specimens collected in Thailand. The developed colorimetric RT-LAMP assay could amplify the target gene and enabled visual interpretation in 60 min at 65 ?C. No cross-reactivity with six other common human respiratory viruses (influenza A virus subtypes H1 and H3, influenza B virus, respiratory syncytial virus types A and B, and human metapneumovirus) and five other human coronaviruses (MERS-CoV, HKU-1, OC43, 229E and NL63) was observed. The limit of detection was 25 copies per reaction when evaluated with contrived specimens. However, the detection rate at this concentration fell to 95.8% when the incubation time was reduced from 60 to 30 min. The diagnostic performance of the developed RT-LAMP assay was evaluated in 2120 clinical specimens and compared with the commercial qRT-PCR. The results revealed high sensitivity and specificity of 95.74% and 99.95%, respectively. The overall accuracy of the RT-LAMP assay was determined to be 99.86%. In summary, our results indicate that the developed colorimetric RT-LAMP provides a simple, sensitive and reliable approach for the detection of SARS-CoV-2 in clinical samples, implying its beneficial use as a diagnostic platform for COVID-19 screening.