Tags

Type your tag names separated by a space and hit enter

Colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a visual diagnostic platform for the detection of the emerging coronavirus SARS-CoV-2.
Analyst. 2021 Jan 21; 146(2):471-477.A

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.

Authors+Show Affiliations

Zenostic Co., Ltd, Bangkok 10400, Thailand. kawin@zenostic.com.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

33165486

Citation

Nawattanapaiboon, Kawin, et al. "Colorimetric Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) as a Visual Diagnostic Platform for the Detection of the Emerging Coronavirus SARS-CoV-2." The Analyst, vol. 146, no. 2, 2021, pp. 471-477.
Nawattanapaiboon K, Pasomsub E, Prombun P, et al. Colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a visual diagnostic platform for the detection of the emerging coronavirus SARS-CoV-2. Analyst. 2021;146(2):471-477.
Nawattanapaiboon, K., Pasomsub, E., Prombun, P., Wongbunmak, A., Jenjitwanich, A., Mahasupachai, P., Vetcho, P., Chayrach, C., Manatjaroenlap, N., Samphaongern, C., Watthanachockchai, T., Leedorkmai, P., Manopwisedjaroen, S., Akkarawongsapat, R., Thitithanyanont, A., Phanchana, M., Panbangred, W., Chauvatcharin, S., & Srikhirin, T. (2021). Colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a visual diagnostic platform for the detection of the emerging coronavirus SARS-CoV-2. The Analyst, 146(2), 471-477. https://doi.org/10.1039/d0an01775b
Nawattanapaiboon K, et al. Colorimetric Reverse Transcription Loop-mediated Isothermal Amplification (RT-LAMP) as a Visual Diagnostic Platform for the Detection of the Emerging Coronavirus SARS-CoV-2. Analyst. 2021 Jan 21;146(2):471-477. PubMed PMID: 33165486.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a visual diagnostic platform for the detection of the emerging coronavirus SARS-CoV-2. AU - Nawattanapaiboon,Kawin, AU - Pasomsub,Ekawat, AU - Prombun,Photchanathorn, AU - Wongbunmak,Akanit, AU - Jenjitwanich,Akarawit, AU - Mahasupachai,Pantanat, AU - Vetcho,Purichaya, AU - Chayrach,Cholticha, AU - Manatjaroenlap,Natthapon, AU - Samphaongern,Chonchanok, AU - Watthanachockchai,Treewat, AU - Leedorkmai,Phonthanat, AU - Manopwisedjaroen,Suwimon, AU - Akkarawongsapat,Radeekorn, AU - Thitithanyanont,Arunee, AU - Phanchana,Matthew, AU - Panbangred,Watanalai, AU - Chauvatcharin,Somchai, AU - Srikhirin,Toemsak, Y1 - 2020/11/09/ PY - 2020/11/10/pubmed PY - 2021/2/2/medline PY - 2020/11/9/entrez SP - 471 EP - 477 JF - The Analyst JO - Analyst VL - 146 IS - 2 N2 - 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. SN - 1364-5528 UR - https://www.unboundmedicine.com/medline/citation/33165486/Colorimetric_reverse_transcription_loop_mediated_isothermal_amplification__RT_LAMP__as_a_visual_diagnostic_platform_for_the_detection_of_the_emerging_coronavirus_SARS_CoV_2_ L2 - https://doi.org/10.1039/d0an01775b DB - PRIME DP - Unbound Medicine ER -