Reverse transcription loop‑mediated isothermal amplification has a high performance in the detection of SARS‑CoV‑2 in saliva samples and nasal swabs from asymptomatic and symptomatic individuals

Reyes-Morales,Rodolfo; Segundo-Ibañez,Patricia; Flores-de Los Ángeles,César; Vizcarra-Ramos,David; Ibañez-Galeana,Damián Iñaki; Salas-Cuevas,Gabriela; Olvera-Serrano,Ángel; Pérez-Silva,Nancy Bibiana; Rocha-Rocha,Valeria Magali; El-Kassi,Elie Girgis; Escobedo-Straffon,Jorge; Contreras-Mioni,Laura; Rosas-Díaz,Marisol; Lopez-Martinez,Karla María; Arias-Matus,Carlos Eduardo; Bautista-Rodriguez,Elizabeth; Nolasco-Quiroga,Manuel

doi:10.3892/etm.2023.12097

Reverse transcription loop‑mediated isothermal amplification has a high performance in the detection of SARS‑CoV‑2 in saliva samples and nasal swabs from asymptomatic and symptomatic individuals

Authors:
- Rodolfo Reyes-Morales
- Patricia Segundo-Ibañez
- César Flores-de Los Ángeles
- David Vizcarra-Ramos
- Damián Iñaki Ibañez-Galeana
- Gabriela Salas-Cuevas
- Ángel Olvera-Serrano
- Nancy Bibiana Pérez-Silva
- Valeria Magali Rocha-Rocha
- Elie Girgis El-Kassi
- Jorge Escobedo-Straffon
- Laura Contreras-Mioni
- Marisol Rosas-Díaz
- Karla María Lopez-Martinez
- Carlos Eduardo Arias-Matus
- Elizabeth Bautista-Rodriguez
- Manuel Nolasco-Quiroga
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Affiliations: Laboratory of Medical and Pharmaceutical Biotechnology, Biotechnology Faculty, Puebla State Popular Autonomous University, Puebla 72410, Mexico, Molecular Biology Laboratory, Biotechnology Department, Interamerican University, Puebla 72828, Mexico, Molecular Diagnostic Laboratory, Biotechnology Faculty, Puebla State Popular Autonomous University, Puebla 72410, Mexico, COVID Area of Hospital Clinic Huauchinango, Institute of Social Security and Services for State Workers, Huauchinango, Puebla 73160, Mexico, Biological Science Department, Biotechnology Faculty, Puebla State Popular Autonomous University, Puebla 72410, Mexico, Molecular Biology Laboratory, Multidisciplinary Academic Unit Reynosa-Aztlan Reynosa, Autonomous University of Tamaulipas, Tamaulipas 88740, Mexico
Published online on: July 6, 2023 https://doi.org/10.3892/etm.2023.12097
Article Number: 398
Copyright: © Reyes-Morales et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The detection of coronavirus disease 2019 cases represents a significant challenge at the epidemiological level. Limitations exist in effectively detecting asymptomatic cases, achieving good follow‑up in hospitals without the infrastructure for reverse transcription‑quantitative PCR (RT‑qPCR) or in difficult‑to‑access areas and developing methods with the need for less invasive sampling procedures. Therefore, the present study evaluated the performance of the direct reverse transcription loop‑mediated isothermal amplification (RT‑LAMP) test for detecting severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) in the saliva and nasal samples of asymptomatic individuals belonging to the university population. In addition, this test was also assessed for effectiveness in symptomatic individuals referred from a hospital with poor infrastructure in molecular biology and located outside the urban area. The RT‑LAMP assay was compared with the results obtained from the RT‑qPCR nasopharyngeal swab test, where the diagnosis was confirmed by lateral flow immunoassay test for rapid antigen detection. A total of 128 samples were analyzed, of which 43% were symptomatic positive individuals, 25% were asymptomatic positive individuals and 32% were SARS‑CoV2‑negative control individuals. Among positive individuals, no differences were found between the Cq values determined by RT‑qPCR. A sensitivity of 96.5% and a specificity of 97.6% was reported for the detection of SARS‑CoV‑2 in symptomatic individuals by salivary and nasal RT‑LAMP, as well as a sensitivity of 100% and a specificity of 97.6% for the detection of SARS‑CoV‑2 in asymptomatic individuals. These findings indicated that performance of the direct RT‑LAMP test using saliva and nasal samples has high sensitivity and specificity, which in turn suggest that it is a viable and reliable alternative for use in epidemiological monitoring.

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