Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers

Barboza, Victor dos Santos; Domingues, William Borges; Souza, Thobias Toniolo de; Collares, Tiago; Seixas, Fabiana Kömmling; Pacheco, Bruna Silveira; Sousa, Fernanda Severo Sabedra; Oliveira, Thaís Larré; Lima, Marcelo de; Pereira, Cláudio M. P.; Spilki, Fernando Rosado; Giongo, Janice Luehring; Vaucher, Rodrigo de Almeida

doi:10.1016/j.jcvp.2023.100134

Cited by 3 publications

(2 citation statements)

References 53 publications

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“…Portable devices can considerably reduce the danger of virus transmission, which made CXRs the favored method of screening during the pandemic; however, the shortage of competent radiologists to interpret imaging data was a significant barrier to CXR screening at the time [42]. A recent virology investigation found that the reverse transcription-loop-mediated isothermal amplification assay was an effective diagnostic tool for use in healthcare personnel [43].…”

Section: Diagnostic Tools and Tracing Strategiesmentioning

confidence: 99%

COVID‐19 retreats and world recovers: A silver lining in the dark cloud

Bisen

Agrawal

Sanap

et al. 2023

Health Care Science

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The coronavirus disease (COVID‐19), which the World Health Organization classified as the Sixth Public Health Emergency Of International Concern (PHEIC) on January 30, 2020, is no longer a PHEIC. Millions were affected due to unawareness. The increase in fatalities and shortage of medicine was the first outrage of COVID‐19. As per the Johns Hopkins COVID‐19 resource center database, it was observed that the disease has spread dynamically across 200+ nations worldwide affecting more than 600 million people from 2019 to 2023, and over thousands of people were victimized regularly at a 2% mortality rate (approx.). In the midway, the mutant variants of concern like omicron, and delta have also created havoc and caused significant impact on public health, global economy, and lifestyle. Since 2019, 3 years now passed and the dynamic disease statistics seem decelerated; moreover, the prevalence of COVID‐19 is also fading. The Johns Hopkins resource center has also stopped recording the data of the global pandemic recently from March 10, 2023. Hence, based on the facts, we are presenting a concise report on the pandemic from 2019 to 2023, which includes a brief discussion of the global pandemic. We have highlighted global epidemiology, emphasizing the Indian COVID scenario, vaccination across the globe, and the psychosocial and geopolitical consequences of COVID‐19 with a brief background to pathology, clinical management, and the worldwide response against triage. A lot has changed and still needs to change after three tough years of COVID‐19. Even though science has progressed and advanced research in medicine is pointing toward future generations, there is no standard care supplied for COVID‐19‐like calamities. COVID‐19 cases might have declined but its influence on the society is still stagnant. This COVID experience has taught us that, despite our bleak beginnings, there is always hope for the future and that we must act with foresight to improve things for future generations.

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Section: Diagnostic Tools and Tracing Strategiesmentioning

confidence: 99%

COVID‐19 retreats and world recovers: A silver lining in the dark cloud

Bisen

Agrawal

Sanap

et al. 2023

Health Care Science

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“…Our research team previously investigated the biotechnological potential of subantarctic algae, such as lipidomics, and, more recently, using κ-carrageenan as a biodegradable film to activate natural oils with antimicrobial protection [ 21 , 22 ]. In addition, new and pioneering approaches to remove SARS-CoV-2 from water and sewage have been developed, along with alternative methods of diagnosing this virus in healthcare workers [ 2 , 6 , 23 , 24 ]. In this study, we aimed to extract and characterize carrageenan from the Chilean macroalgae G. skottsbergii for use in molecular methods as a binding probe for the SARS-CoV-2 virus.…”

Section: Introductionmentioning

confidence: 99%

Carrageenan from Gigartina skottsbergii: A Novel Molecular Probe to Detect SARS-CoV-2

et al. 2023

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The COVID-19 pandemic has caused an unprecedented health and economic crisis, highlighting the importance of developing new molecular tools to monitor and detect SARS-CoV-2. Hence, this study proposed to employ the carrageenan extracted from Gigartina skottsbergii algae as a probe for SARS-CoV-2 virus binding capacity and potential use in molecular methods. G. skottsbergii specimens were collected in the Chilean subantarctic ecoregion, and the carrageenan was extracted —using a modified version of Webber’s method—, characterized, and quantified. After 24 h of incubation with an inactivated viral suspension, the carrageenan’s capacity to bind SARS-CoV-2 was tested. The probe-bound viral RNA was quantified using the reverse transcription and reverse transcription loop-mediated isothermal amplification (RT-LAMP) methods. Our findings showed that carrageenan extraction from seaweed has a similar spectrum to commercial carrageenan, achieving an excellent proportion of binding to SARS-CoV-2, with a yield of 8.3%. Viral RNA was also detected in the RT-LAMP assay. This study shows, for the first time, the binding capacity of carrageenan extracted from G. skottsbergii, which proved to be a low-cost and highly efficient method of binding to SARS-CoV-2 viral particles.

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Sample Treatment with Trypsin for RT-LAMP COVID-19 Diagnosis

García-Sorribes,

Lara-Hernández,

Manzano-Blasco

et al. 2023

Biology

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The SARS-CoV-2 coronavirus is responsible for the COVID-19 pandemic resulting in a global health emergency. Given its rapid spread and high number of infected individuals, a diagnostic tool for a rapid, simple, and cost-effective detection was essential. In this work, we developed a COVID-19 diagnostic test, that incorporates a human internal control, based on the Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP). When working with synthetic SARS-CoV-2 RNA, the optimized RT-LAMP assay has a sensitivity of 10 viral copies and can be detected by fluorescence in less than 15 min or by the naked eye in 25 min using colorimetric RT-LAMP. To avoid the RNA extraction step, a pre-treatment of the sample was optimized. Subsequently, a validation was performed on 268 trypsin treated samples (including nasopharyngeal, buccal, and nasal exudates) and amplified with colorimetric RT-LAMP to evaluate its sensitivity and specificity in comparison with RT-qPCR of extracted samples. The validation results showed a sensitivity and specificity of 100% for samples with Ct ≤ 30. The rapid, simple, and inexpensive RT-LAMP SARS-CoV-2 extraction-free procedure developed may be an alternative test that could be applied for the detection of SARS-CoV-2 or adapted to detect other viruses present in saliva or nasopharyngeal samples with higher sensitivity and specificity of the antibody test.

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Reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay as a rapid molecular diagnostic tool for COVID-19 in healthcare workers

Cited by 3 publications

References 53 publications

COVID‐19 retreats and world recovers: A silver lining in the dark cloud

COVID‐19 retreats and world recovers: A silver lining in the dark cloud

Carrageenan from Gigartina skottsbergii: A Novel Molecular Probe to Detect SARS-CoV-2

Sample Treatment with Trypsin for RT-LAMP COVID-19 Diagnosis

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