SARS-CoV-2 RT-LAMP检测的标准化和性能:一种可靠、廉价和可替代的诊断方法。

IF 1.9 4区 生物学 Q3 MICROBIOLOGY
Rachel Cruz Alves, José Henrique Franscisco Roma, Bruno Moreira Carneiro, Juliana Helena Chavez Pavoni, Renata Dezengrini Slhessarenko
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引用次数: 0

摘要

SARS-CoV-2的出现标志着COVID-19大流行的开始,这对全球公共卫生构成了挑战。采用不同检测方法进行大规模检测是控制病毒传播的一项基本战略,特别是在低收入地区。本研究旨在标准化和验证RT-LAMP作为SARS-CoV-2检测的替代工具。对不同的引物进行了硅质和体外评价,并选择N2靶标进入验证阶段。从临床样品中提取的RNA用BST 3.0和RT酶对RT-LAMP荧光法进行标准化,用WarmStart®colorimetric LAMP Master Mix进行比色分析。临床样本也进行了wonfo 2019-nCoV抗原检测。所提出的方案在高病毒载量样品(CT≤30)中显示出强大的诊断准确性,荧光法检测灵敏度为98.25%,特异性为90.91%,比色法检测灵敏度为92.98%,特异性为100%。考虑到两种可视化技术,荧光技术得到了更一致的结果。相比之下,快速抗原检测的表现较差,在CT评分≤30的样本中,敏感性为82.46%,特异性为100%。简单紫外光源的RT-LAMP可视化在活动性感染患者样品中显示出较高的灵敏度。更快、最简单的执行、成本效益和对结果的可理解解释是改进这一诊断工具作为COVID-19主动传播的潜在筛查工具的有利因素,特别是在财政资源有限的地区。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Standardization and performance of SARS-CoV-2 RT-LAMP detection: a reliable, inexpensive, and alternative diagnostic assay.

The emergence of SARS-CoV-2 marked the onset of the COVID-19 pandemic, which has challenged public health worldwide. Mass testing performed by different assays represents an essential strategy to control virus spread, especially in low-income regions. This study aimed to standardize and validate RT-LAMP as an alternative tool for SARS-CoV-2 detection. Different sets of primers were assessed in silico and in vitro, and the N2 target was selected for the validation stage. The extracted RNA from the clinical samples was used to standardize the RT-LAMP fluorometric assay with BST 3.0 and RT enzymes, and the colorimetric assay was performed with WarmStart® Colorimetric LAMP Master Mix. Clinical samples were also subjected to the Wondfo 2019-nCoV antigen test. The proposed protocols showed robust diagnostic accuracy in high-viral-load samples (CT ≤ 30), 98.25% sensitivity and 90.91% specificity for fluorometric assay, and 92.98% sensitivity and 100% specificity for colorimetric assay. Considering the two visualization techniques, the fluorometric technique yielded more accordant results. In contrast, the rapid antigen test presented a lower performance, with 82.46% sensitivity and 100% specificity in samples with a CT score ≤ 30. RT-LAMP visualization with a simple ultraviolet light source showed high sensitivity in active infection patient samples. The faster and simplest execution, cost-effectiveness, and accessible interpretation of the results are favorable points for improving this diagnostic tool as a potential screening tool for active transmission of COVID-19, especially in regions with limited financial resources.

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来源期刊
Brazilian Journal of Microbiology
Brazilian Journal of Microbiology 生物-微生物学
CiteScore
4.10
自引率
4.50%
发文量
216
审稿时长
1.0 months
期刊介绍: The Brazilian Journal of Microbiology is an international peer reviewed journal that covers a wide-range of research on fundamental and applied aspects of microbiology. The journal considers for publication original research articles, short communications, reviews, and letters to the editor, that may be submitted to the following sections: Biotechnology and Industrial Microbiology, Food Microbiology, Bacterial and Fungal Pathogenesis, Clinical Microbiology, Environmental Microbiology, Veterinary Microbiology, Fungal and Bacterial Physiology, Bacterial, Fungal and Virus Molecular Biology, Education in Microbiology. For more details on each section, please check out the instructions for authors. The journal is the official publication of the Brazilian Society of Microbiology and currently publishes 4 issues per year.
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