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
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.
Exp Ther Med. 2023 Jul 6;26(2):398. doi: 10.3892/etm.2023.12097. eCollection 2023 Aug.
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.
2019冠状病毒病病例的检测在流行病学层面是一项重大挑战。在有效检测无症状病例、在没有逆转录定量聚合酶链反应(RT-qPCR)基础设施的医院或难以进入的地区实现良好随访以及开发需要侵入性较小采样程序的方法方面存在局限性。因此,本研究评估了直接逆转录环介导等温扩增(RT-LAMP)试验在检测大学人群无症状个体唾液和鼻腔样本中严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的性能。此外,还评估了该试验在从分子生物学基础设施较差且位于市区外的医院转诊的有症状个体中的有效性。将RT-LAMP检测结果与RT-qPCR鼻咽拭子检测结果进行比较,后者通过侧向流免疫分析快速抗原检测来确诊。共分析了128份样本,其中43%为有症状阳性个体,25%为无症状阳性个体,32%为SARS-CoV-2阴性对照个体。在阳性个体中,RT-qPCR测定的Cq值之间未发现差异。唾液和鼻腔RT-LAMP检测有症状个体中SARS-CoV-2的灵敏度为96.5%,特异性为97.6%,检测无症状个体中SARS-CoV-2的灵敏度为100%,特异性为97.6%。这些发现表明,使用唾液和鼻腔样本的直接RT-LAMP检测具有高灵敏度和特异性,这反过来表明它是流行病学监测中一种可行且可靠的替代方法。