Tags

Type your tag names separated by a space and hit enter

Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2.
J Virol Methods. 2020 12; 286:113972.JV

Abstract

A novel reverse-transcriptase loop mediated amplification (RT-LAMP) method targeting genes encoding the Spike (S) protein and RNA-dependent RNA polymerase (RdRP) of SARS-CoV-2 has been developed. The LAMP assay achieves a comparable limit of detection (25-50 copies per reaction) to commonly used RT-PCR protocols using clinical samples quantified by digital droplet PCR. Precision, cross-reactivity, inclusivity, and limit of detection studies were performed according to regulatory standards. Clinical validation of dual-target RT-LAMP (S and RdRP gene) achieved a PPA of 98.48 % (95 % CI 91.84%-99.96%) and NPA 100.00 % (95 % CI 93.84%-100.00%) based on the E gene and N2 gene reference RT-PCR methods. The method has implications for development of point of care technology using isothermal amplification.

Authors+Show Affiliations

Department of Microbiology, Immunology, and Infectious Diseases, University of Calgary, AB, Canada.Department of Microbiology, Immunology, and Infectious Diseases, University of Calgary, AB, Canada.Department of Microbiology, Immunology, and Infectious Diseases, University of Calgary, AB, Canada; Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.Department of Microbiology, Immunology, and Infectious Diseases, University of Calgary, AB, Canada.Alberta Public Health Laboratory, Alberta Precision Laboratory, Calgary, AB, Canada.Alberta Public Health Laboratory, Alberta Precision Laboratory, Calgary, AB, Canada; Clinical Section of Microbiology, Alberta Precision Laboratories, Calgary, AB, Canada.Clinical Section of Microbiology, Alberta Precision Laboratories, Calgary, AB, Canada; Department Pathology and Laboratory Medicine, University of Calgary, Calgary, AB, Canada.Clinical Section of Microbiology, Alberta Precision Laboratories, Calgary, AB, Canada; Department Pathology and Laboratory Medicine, University of Calgary, Calgary, AB, Canada.Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, Canada.Illucidx Inc., Calgary, AB, Canada.Department Pathology and Laboratory Medicine, University of California, San Francisco, USA.Department Pathology and Laboratory Medicine, University of California, San Francisco, USA.Department of Laboratory Medicine, University of Washington, Seattle, WA, USA.Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA.Department of Microbiology, Immunology, and Infectious Diseases, University of Calgary, AB, Canada; Clinical Section of Microbiology, Alberta Precision Laboratories, Calgary, AB, Canada; Department Pathology and Laboratory Medicine, University of Calgary, Calgary, AB, Canada; Department of Medicine, University of Calgary, Calgary, AB, Canada. Electronic address: drpillai@ucalgary.ca.

Pub Type(s)

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

Language

eng

PubMed ID

32941977

Citation

Mohon, Abu Naser, et al. "Optimization and Clinical Validation of Dual-target RT-LAMP for SARS-CoV-2." Journal of Virological Methods, vol. 286, 2020, p. 113972.
Mohon AN, Oberding L, Hundt J, et al. Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2. J Virol Methods. 2020;286:113972.
Mohon, A. N., Oberding, L., Hundt, J., van Marle, G., Pabbaraju, K., Berenger, B. M., Lisboa, L., Griener, T., Czub, M., Doolan, C., Servellita, V., Chiu, C. Y., Greninger, A. L., Jerome, K. R., & Pillai, D. R. (2020). Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2. Journal of Virological Methods, 286, 113972. https://doi.org/10.1016/j.jviromet.2020.113972
Mohon AN, et al. Optimization and Clinical Validation of Dual-target RT-LAMP for SARS-CoV-2. J Virol Methods. 2020;286:113972. PubMed PMID: 32941977.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Optimization and clinical validation of dual-target RT-LAMP for SARS-CoV-2. AU - Mohon,Abu Naser, AU - Oberding,Lisa, AU - Hundt,Jana, AU - van Marle,Guido, AU - Pabbaraju,Kanti, AU - Berenger,Byron M, AU - Lisboa,Luiz, AU - Griener,Thomas, AU - Czub,Markus, AU - Doolan,Cody, AU - Servellita,Venice, AU - Chiu,Charles Y, AU - Greninger,Alexander L, AU - Jerome,Keith R, AU - Pillai,Dylan R, Y1 - 2020/09/15/ PY - 2020/07/06/received PY - 2020/09/04/revised PY - 2020/09/11/accepted PY - 2020/9/18/pubmed PY - 2020/12/1/medline PY - 2020/9/17/entrez KW - Assay KW - Covid KW - Diagnostics KW - LAMP KW - Molecular KW - PCR KW - SARS-CoV-2 KW - Validation SP - 113972 EP - 113972 JF - Journal of virological methods JO - J Virol Methods VL - 286 N2 - A novel reverse-transcriptase loop mediated amplification (RT-LAMP) method targeting genes encoding the Spike (S) protein and RNA-dependent RNA polymerase (RdRP) of SARS-CoV-2 has been developed. The LAMP assay achieves a comparable limit of detection (25-50 copies per reaction) to commonly used RT-PCR protocols using clinical samples quantified by digital droplet PCR. Precision, cross-reactivity, inclusivity, and limit of detection studies were performed according to regulatory standards. Clinical validation of dual-target RT-LAMP (S and RdRP gene) achieved a PPA of 98.48 % (95 % CI 91.84%-99.96%) and NPA 100.00 % (95 % CI 93.84%-100.00%) based on the E gene and N2 gene reference RT-PCR methods. The method has implications for development of point of care technology using isothermal amplification. SN - 1879-0984 UR - https://www.unboundmedicine.com/medline/citation/32941977/Optimization_and_clinical_validation_of_dual_target_RT_LAMP_for_SARS_CoV_2_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0166-0934(20)30224-X DB - PRIME DP - Unbound Medicine ER -