Detection of SARS-CoV-2 RNA in Urine by RT-LAMP: A Very Rare Finding

Diego, Juan García-Bernalt; Fernández‐Soto, Pedro; Muñoz‐Bellido, Juan Luis; Febrer‐Sendra, Begoña; Crego-Vicente, Beatriz; Carbonell, Cristina; López-Bernús, Amparo; Marcos, Miguel; Belhassen‐García, Moncef; Muro, Antonio

doi:10.3390/jcm11010158

Cited by 5 publications

(1 citation statement)

References 35 publications

(46 reference statements)

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“…This technique was used to test the urine of 300 patients divided into three groups: RT-qPCR COVID-19 positive using a nasopharyngeal swab (n = 100), COVID-19 negative (n = 100), and COVID-19 positive with disease recovery (n = 100). The results showed a trace amount of SARS-CoV-2 RNA, with only 4% positivity found in the positive group [ 19 ]. Other studies continued to prove the reliability of LAMP assay in detecting the SARS-CoV-2 virus using different sampling locations.…”

Section: Introductionmentioning

confidence: 99%

Development and Validation of Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) as a Simple and Rapid Diagnostic Tool for SARS-CoV-2 Detection

et al. 2022

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Since the COVID-19 pandemic outbreak in the world, many countries have searched for quick diagnostic tools to detect the virus. There are many ways to design diagnostic assays; however, each may have its limitations. A quick, sensitive, specific, and simple approach is essential for highly rapidly transmitted infections, such as SARS-CoV-2. This study aimed to develop a rapid and cost-effective diagnostic tool using a one-step Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) approach. The results were observed using the naked eye within 30–60 min using turbidity or colorimetric analysis. The sensitivity, specificity, and lowest limit of detection (LoD) for SARS-CoV-2 RNA against the RT-LAMP assay were assessed. This assay was also verified and validated against commercial quantitative RT-PCR used by health authorities in Saudi Arabia. Furthermore, a quick and direct sampling from the saliva, or buccal cavity, was applied after simple modification, using proteinase K and heating at 98 °C for 5 min to avoid routine RNA extraction. This rapid single-tube diagnostic tool detected COVID-19 with an accuracy rate of 95% for both genes ( ORF1a and N ) and an LoD for the ORF1a and N genes as 39 and 25 copies/reaction, respectively. It can be potentially used as a high-throughput national screening for different respiratory-based infections within the Middle East region, such as the MERS virus or major zoonotic pathogens such as Mycobacterium paratuberculosis and Brucella spp., particularly in remote and rural areas where lab equipment is limited.

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Section: Introductionmentioning

confidence: 99%

Development and Validation of Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) as a Simple and Rapid Diagnostic Tool for SARS-CoV-2 Detection

et al. 2022

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A Clinical Assessment of the RT-LAMP-Based Colorimetric Diagnostic Method for COVID-19 with Improved Primers Sets

Sarkar,

Balasubramanian,

Velugonda

et al. 2023

Appl Biochem Microbiol

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First field and laboratory evaluation of LAMP assay for malaria diagnosis in Cubal, Angola

Febrer-Sendra,

Crego-Vicente,

Nindia

et al. 2023

Parasites Vectors

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Background Malaria is a globally distributed infectious disease. According to the World Health Organization, Angola is one of the six countries that account for over half the global malaria burden in terms of both malaria cases and deaths. Diagnosis of malaria still depends on microscopic examination of thin and thick blood smears and rapid diagnostic tests (RDTs), which often lack analytical and clinical sensitivity. Molecular methods could overcome these disadvantages. The aim of this study was to evaluate, for the first time to our knowledge, the performance of a loop-mediated isothermal amplification (LAMP) for the diagnosis of malaria in an endemic area in Cubal, Angola, and to assess the reproducibility at a reference laboratory. Methods A total of 200 blood samples from patients attended at Hospital Nossa Senhora da Paz, Cubal, Angola, were analysed for Plasmodium spp. detection by microscopy, RDTs, and LAMP. LAMP assay was easily performed in a portable heating block, and the results were visualized by a simple colour change. Subsequently, the samples were sent to a reference laboratory in Spain to be reanalysed by the same colorimetric LAMP assay and also in real-time LAMP format. Results In field tests, a total of 67/200 (33.5%) blood samples were microscopy-positive for Plasmodium spp., 98/200 RDT positive, and 112/200 (56%) LAMP positive. Using microscopy as reference standard, field LAMP detected more microscopy-positive samples than RDTs (66/67; 98% vs. 62/67; 92.5%). When samples were reanalysed at a reference laboratory in Spain using both colorimetric and real-time assays, the overall reproducibility achieved 84.5%. Conclusions This is the first study to our knowledge in which LAMP has been clinically evaluated on blood samples in a resource-poor malaria-endemic area. The colorimetric LAMP proved to be more sensitive than microscopy and RDTs for malaria diagnosis in field conditions. Furthermore, LAMP showed an acceptable level of reproducibility in a reference laboratory. The possibility to use LAMP in a real-time format in a portable device reinforces the reliability of the assay for molecular diagnosis of malaria in resource-poor laboratories in endemic areas. Graphical Abstract

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Detection of SARS-CoV-2 RNA in Urine by RT-LAMP: A Very Rare Finding

Cited by 5 publications

References 35 publications

Development and Validation of Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) as a Simple and Rapid Diagnostic Tool for SARS-CoV-2 Detection

Development and Validation of Reverse Transcriptase Loop-Mediated Isothermal Amplification (RT-LAMP) as a Simple and Rapid Diagnostic Tool for SARS-CoV-2 Detection

A Clinical Assessment of the RT-LAMP-Based Colorimetric Diagnostic Method for COVID-19 with Improved Primers Sets

First field and laboratory evaluation of LAMP assay for malaria diagnosis in Cubal, Angola

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