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Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2.
PLoS One. 2021; 16(3):e0248042.Plos

Abstract

A newly identified coronavirus, designated as severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), has spread rapidly from its epicenter in China to more than 150 countries across six continents. In this study, we have designed three reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primer sets to detect the RNA-dependent RNA polymerase (RdRP), Envelope (E) and Nucleocapsid protein (N) genes of SARS CoV-2. For one tube reaction, the detection limits for five combination SARS CoV-2 LAMP primer sets (RdRP/E, RdRP/N, E/N, RdRP/E/N and RdRP/N/Internal control (actin beta)) were evaluated with a clinical nasopharyngeal swab sample. Among the five combination, the RdRP/E and RdRP/N/IC multiplex LAMP assays showed low detection limits. The sensitivity and specificity of the RT-LAMP assay were evaluated and compared to that of the widely used Allplex™ 2019-nCoV Assay (Seegene, Inc., Seoul, South Korea) and PowerChek™ 2019-nCoV Real-time PCR kit (Kogenebiotech, Seoul, South Korea) for 130 clinical samples from 91 SARS CoV-2 patients and 162 NP specimens from individuals with (72) and without (90) viral respiratory infections. The multiplex RdRP (FAM)/N (CY5)/IC (Hex) RT-LAMP assay showed comparable sensitivities (RdRP: 93.85%, N: 94.62% and RdRP/N: 96.92%) to that of the Allplex™ 2019-nCoV Assay (100%) and superior to those of PowerChek™ 2019-nCoV Real-time PCR kit (RdRP: 92.31%, E: 93.85% and RdRP/E: 95.38%).

Authors+Show Affiliations

Emergency Medicine, College of Medicine, Korea University Guro Hospital, Seoul, Republic of Korea.Departments of Laboratory Medicine, College of Medicine, Korea University Guro Hospital, Seoul, Republic of Korea.Departments of Laboratory Medicine, College of Medicine, Korea University Guro Hospital, Seoul, Republic of Korea.Departments of Laboratory Medicine, College of Medicine, Korea University Guro Hospital, Seoul, Republic of Korea.Emergency Medicine, College of Medicine, Korea University Guro Hospital, Seoul, Republic of Korea.Emergency Medicine, College of Medicine, Korea University Guro Hospital, Seoul, Republic of Korea.Pacific Center for Emerging Infectious Diseases Research, John A. Burns School of Medicine, University of Hawaii at Manoa, Honolulu, HI, United States of America.Department of Laboratory Medicine, Kangwon National University, School of Medicine, Kangwondo, Republic of Korea.Department of Laboratory Medicine, Dankook University College of Medicine, Cheonan, Korea.Department of Laboratory Medicine, Dongsan Medical Center, Keimyung University, Daegu, Korea.Departments of Laboratory Medicine, College of Medicine, Korea University Guro Hospital, Seoul, Republic of Korea.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

33657176

Citation

Jang, Woong Sik, et al. "Development of a Multiplex Loop-Mediated Isothermal Amplification (LAMP) Assay for On-site Diagnosis of SARS CoV-2." PloS One, vol. 16, no. 3, 2021, pp. e0248042.
Jang WS, Lim DH, Yoon J, et al. Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2. PLoS One. 2021;16(3):e0248042.
Jang, W. S., Lim, D. H., Yoon, J., Kim, A., Lim, M., Nam, J., Yanagihara, R., Ryu, S. W., Jung, B. K., Ryoo, N. H., & Lim, C. S. (2021). Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2. PloS One, 16(3), e0248042. https://doi.org/10.1371/journal.pone.0248042
Jang WS, et al. Development of a Multiplex Loop-Mediated Isothermal Amplification (LAMP) Assay for On-site Diagnosis of SARS CoV-2. PLoS One. 2021;16(3):e0248042. PubMed PMID: 33657176.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Development of a multiplex Loop-Mediated Isothermal Amplification (LAMP) assay for on-site diagnosis of SARS CoV-2. AU - Jang,Woong Sik, AU - Lim,Da Hye, AU - Yoon,Jung, AU - Kim,Ahran, AU - Lim,Minsup, AU - Nam,Jeonghun, AU - Yanagihara,Richard, AU - Ryu,Sook-Won, AU - Jung,Bo Kyeung, AU - Ryoo,Nam-Hee, AU - Lim,Chae Seung, Y1 - 2021/03/03/ PY - 2020/12/27/received PY - 2021/02/18/accepted PY - 2021/3/3/entrez PY - 2021/3/4/pubmed PY - 2021/4/1/medline SP - e0248042 EP - e0248042 JF - PloS one JO - PLoS One VL - 16 IS - 3 N2 - A newly identified coronavirus, designated as severe acute respiratory syndrome coronavirus 2 (SARS CoV-2), has spread rapidly from its epicenter in China to more than 150 countries across six continents. In this study, we have designed three reverse-transcription loop-mediated isothermal amplification (RT-LAMP) primer sets to detect the RNA-dependent RNA polymerase (RdRP), Envelope (E) and Nucleocapsid protein (N) genes of SARS CoV-2. For one tube reaction, the detection limits for five combination SARS CoV-2 LAMP primer sets (RdRP/E, RdRP/N, E/N, RdRP/E/N and RdRP/N/Internal control (actin beta)) were evaluated with a clinical nasopharyngeal swab sample. Among the five combination, the RdRP/E and RdRP/N/IC multiplex LAMP assays showed low detection limits. The sensitivity and specificity of the RT-LAMP assay were evaluated and compared to that of the widely used Allplex™ 2019-nCoV Assay (Seegene, Inc., Seoul, South Korea) and PowerChek™ 2019-nCoV Real-time PCR kit (Kogenebiotech, Seoul, South Korea) for 130 clinical samples from 91 SARS CoV-2 patients and 162 NP specimens from individuals with (72) and without (90) viral respiratory infections. The multiplex RdRP (FAM)/N (CY5)/IC (Hex) RT-LAMP assay showed comparable sensitivities (RdRP: 93.85%, N: 94.62% and RdRP/N: 96.92%) to that of the Allplex™ 2019-nCoV Assay (100%) and superior to those of PowerChek™ 2019-nCoV Real-time PCR kit (RdRP: 92.31%, E: 93.85% and RdRP/E: 95.38%). SN - 1932-6203 UR - https://www.unboundmedicine.com/medline/citation/33657176/Development_of_a_multiplex_Loop_Mediated_Isothermal_Amplification__LAMP__assay_for_on_site_diagnosis_of_SARS_CoV_2_ L2 - https://dx.plos.org/10.1371/journal.pone.0248042 DB - PRIME DP - Unbound Medicine ER -