COVID-19的护理点检测:在SARS-CoV-2大流行期间开发和验证简单的两步分子诊断法。

IF 2.5 4区 医学 Q2 PARASITOLOGY
Memorias do Instituto Oswaldo Cruz Pub Date : 2024-10-04 eCollection Date: 2024-01-01 DOI:10.1590/0074-02760230236
Andre Akira Gonzaga Yoshikawa, Sabrina Fernandes Cardoso, Lívia Budziarek Eslabão, Iara Carolini Pinheiro, Priscila Valverde, Gisele Caminha, Oscar Bruna Romero, Leandro Medeiros, Luísa Damazio Pitaluga Rona, André Nóbrega Pitaluga
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引用次数: 0

摘要

背景:在冠状病毒疾病 19(COVID-19)大流行期间,由于使用反转录实时聚合酶链反应(RT-qPCR)的黄金标准诊断程序在集中模式下进行大规模检测的局限性,对普通人群进行诊断检测具有挑战性,特别是在低资源地区:为解决这一问题,我们开发了一种针对 COVID-19 的护理点(PoC)诊断方案,提供快速、可靠且经济实惠的检测,尤其适用于中低端发展地区:方法:PoC 诊断过程将一种简单的纸质 RNA 提取方法与比色反转录环介导等温扩增(RT-LAMP)检测相结合,该方法安装在一个 3D 打印的塑料装置中。在巴西圣卡塔琳娜州卫生部长的协助下,2020 年至 2021 年期间对鼻咽/咽拭子(NOS)和唾液样本进行了检测:所制定的诊断方案显示,从 1,348 份临床分析样本中,严重急性呼吸系统综合征冠状病毒 2(SARS-CoV-2)的检测限为 9,900 个拷贝,总体诊断特异性为 98%。对 NOS 样本的诊断灵敏度为 95%,对清晨唾液的诊断灵敏度为 85%,对任意唾液的诊断灵敏度为 69%:主要结论:总之,所开发的装置成功地从咽拭子和唾液临床样本中提取了 SARS-CoV-2 病毒 RNA。当与比色 RT-LAMP 结合使用时,它能在 45 分钟内利用最少的资源得出结果,从而提供了一种适用于大规模采样的诊断试剂盒方案。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Point-of-care testing for COVID-19: a simple two-step molecular diagnostic development and validation during the SARS-CoV-2 pandemic.

Background: During the coronavirus disease 19 (COVID-19) pandemic, diagnostic testing of the general population proved challenging due to limitations of the gold-standard diagnostic procedure using reverse transcription real-time polymerase chain reaction (RT-qPCR) for large-scale testing on the centralised model, especially in low-resource areas.

Objectives: To address this, a point-of-care (PoC) diagnostic protocol for COVID-19 was developed, providing fast, reliable, and affordable testing, particularly for low-mid develop areas.

Methods: The PoC diagnostic process combines a simple paper-based RNA extraction method housed within a 3D-printed plastic device with a colorimetric reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay. Nasopharyngeal/oropharyngeal swabs (NOS) and saliva samples were tested between 2020 and 2021, with the assistance of Santa Catarina's State Health Secretary, Brazil.

Findings: The developed diagnostic protocol showed a limit of detection of 9,900 copies and an overall diagnostic specificity of 98% for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from 1,348 clinical analysed samples. The diagnostic sensitivity was 95% for NOS samples, 85% for early morning saliva, and 69% for indiscriminate saliva.

Main conclusions: In conclusion, the developed device successfully extracted SARS-CoV-2 viral RNA from swabs and saliva clinical samples. When combined with colorimetric RT-LAMP, it provides results within 45 min using minimal resources, thus delivering a diagnostic kit protocol that is applicable in large-scale sampling.

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来源期刊
CiteScore
5.00
自引率
3.60%
发文量
91
审稿时长
3-8 weeks
期刊介绍: Memórias do Instituto Oswaldo Cruz is a journal specialized in microbes & their vectors causing human infections. This means that we accept manuscripts covering multidisciplinary approaches and findings in the basic aspects of infectious diseases, e.g. basic in research in prokariotes, eukaryotes, and/or virus. Articles must clearly show what is the main question to be answered, the hypothesis raised, and the contribution given by the study. Priority is given to manuscripts reporting novel mechanisms and general findings concerning the biology of human infectious prokariotes, eukariotes or virus. Papers reporting innovative methods for diagnostics or that advance the basic research with these infectious agents are also welcome. It is important to mention what we do not publish: veterinary infectious agents research, taxonomic analysis and re-description of species, epidemiological studies or surveys or case reports and data re-analysis. Manuscripts that fall in these cases or that are considered of low priority by the journal editorial board, will be returned to the author(s) for submission to another journal.
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