scholarly journals Analysis, quantification, and visualization of RT-LAMP technique for detection of COVID-19

Author(s):  
Mohammad Hosein Ghazimoradi ◽  
Maryam Daryani ◽  
Masoud Gharshasbi ◽  
Ehsan Zolghadr ◽  
Ali Khalafizadeh ◽  
...  

Background: SARS-Cov-2 is a new virus that caused an epidemic disease, COVID-19. According to the world health organization, detecting the patients/carriers is by the far the most important action to prevent the pandemic. Recently, the loop-mediated isothermal amplification (LAMP) technique has become more popular due to the easy handling of a one-step kit used for the detection of many diseases than RT-PCR-based techniques. Methods: Herein, we used the RT-LAMP technique so as to detect COVID-19. To this end, 40 paired-samples of patients and healthy people had been collected and tested by RT-PCR for N and E genes of SARS-CoV-2. The RT-LAMP test has been performed on samples for the RdRp gene. The sensitivity and specificity of tests have been determined. Results: The testing results are consistent with the conventional RT-qPCR. Additionally, we also showed that a one-step process without RNA extraction is feasible to achieve RNA amplification directly from a sample. Conclusion: We confirmed that RT-LAMP is a rapid, simple, and sensitive method that can be used as a large-screening method, particularly in regional hospitals with limited access to high-technologies.

2020 ◽  
Author(s):  
Jijoho Michel Agbla ◽  
Mathew D Esona ◽  
Alidéhou Jerrold Agbankpé ◽  
Annick Capo-Chichi ◽  
Rashi Gautam ◽  
...  

Abstract Objective: Rotavirus remains the main causative agent of gastroenteritis in young children in countries that have not yet introduced the vaccine. In Benin, rotavirus vaccine was introduced late December 2019 into the EPI. This study aims to provide pre-vaccination era rotavirus genotyping data in Benin. These data can supplement data from the surveillance system of Ministry of Health of Benin which is supported by the World Health Organization (WHO).Results: Of the 420 diarrheal stool samples, actively collected in southern Benin from July 2016 through November 2018 from children under five years old and suffering from gastroenteritis, 167 (39.8%) samples were rotavirus EIA positive. 186 (44.3%) samples contained amplifiable rotavirus RNA detected by qRT-PCR method and were genotyped using one-step RT-PCR multiplex genotyping method. G1P[8] represents the predominant genotype (32%) followed by the G2P[4] (26%), G3P[6] (16%), G12P[8] (13%) and mixed G and P types (1%). Four samples (2%) could not be assigned both G and P type specificity.


2021 ◽  
Author(s):  
Jijoho Michel Agbla ◽  
Mathew D Esona ◽  
Alidéhou Jerrold Agbankpé ◽  
Annick Capo-Chichi ◽  
Rashi Gautam ◽  
...  

Abstract ObjectiveRotavirus remains the main causative agent of gastroenteritis in young children in countries that have not yet introduced the vaccine. In Benin, rotavirus vaccine was introduced late December 2019 into the EPI. This study aims to provide pre-vaccination era rotavirus genotyping data in Benin. These data can supplement data from the surveillance system of Ministry of Health of Benin which is supported by the World Health Organization (WHO).ResultsOf the 420 diarrheal stool samples, actively collected in southern Benin from July 2016 through November 2018 from children under five years old and suffering from gastroenteritis, 167 (39.8%) samples were rotavirus EIA positive. 186 (44.3%) samples contained amplifiable rotavirus RNA detected by qRT-PCR method and were genotyped using one-step RT-PCR multiplex genotyping method. G1P[8] represents the predominant genotype (32%) followed by the G2P[4] (26%), G3P[6] (16%), G12P[8] (13%) and mixed G and P types (1%). Four samples (2%) could not be assigned both G and P type specificity.


2020 ◽  
Author(s):  
Jijoho Michel Agbla ◽  
Mathew D Esona ◽  
Alidéhou Jerrold Agbankpé ◽  
Annick Capo-Chichi ◽  
Rashi Gautam ◽  
...  

Abstract ObjectiveRotavirus remains the main causative agent of gastroenteritis in young children in countries that have not yet introduced the vaccine. In Benin, rotavirus vaccine was introduced late December 2019 into the EPI. This study aims to provide pre-vaccination era rotavirus genotyping data in Benin. These data can supplement data from the surveillance system of Ministry of Health of Benin which is supported by the World Health Organization (WHO). ResultsOf the 420 diarrheal stool samples, actively collected in southern Benin from July 2016 through November 2018 from children under five years old and suffering from gastroenteritis, 167 (39.8%) samples were rotavirus EIA positive. 186 (44.3%) samples contained amplifiable rotavirus RNA detected by qRT-PCR method and were genotyped using one-step RT-PCR multiplex genotyping method. G1P[8] represents the predominant genotype (32%) followed by the G2P[4] (26%), G3P[6] (16%), G12P[8] (13%) and mixed G and P types (1%). Four samples (2%) could not be assigned both G and P type specificity.


2020 ◽  
Author(s):  
Jijoho Michel Agbla ◽  
Mathew D Esona ◽  
Alidéhou Jerrold Agbankpé ◽  
Annick Capo-Chichi ◽  
Rashi Gautam ◽  
...  

Abstract Objective Rotavirus remains the main causative agent of gastroenteritis in young children in countries that have not yet introduced the vaccine. In Benin, rotavirus vaccine was introduced late December 2019 into the EPI. This study aims to provide pre-vaccination era rotavirus genotyping data in Benin. These data can supplement data from the surveillance system of Ministry of Health of Benin which is supported by the World Health Organization (WHO). Results Of the 420 diarrheal stool samples, actively collected in southern Benin from July 2016 through November 2018 from children under five years old and suffering from gastroenteritis, 167 (39.8%) samples were rotavirus EIA positive. 186 (44.3%) samples contained amplifiable rotavirus RNA detected by qRT-PCR method and were genotyped using one-step RT-PCR multiplex genotyping method. G1P[8] represents the predominant genotype (32%) followed by the G2P[4] (26%), G3P[6] (16%), G12P[8] (13%) and mixed G and P types (1%). Four samples (2%) could not be assigned both G and P type specificity.


Author(s):  
Xue-fei Cai ◽  
Juan Chen ◽  
Jie-li Hu ◽  
Quan-xin Long ◽  
Hai-jun Deng ◽  
...  

AbstractA respiratory illness has been spreading rapidly in China, since its outbreak in Wuhan city, Hubei province in December 2019. The illness was caused by a novel coronavirus, named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Clinical manifestations related to SARS-CoV-2 infection ranged from no symptom to fatal pneumonia. World Health Organization (WHO) named the diseases associated with SARS-CoV-2 infection as COVID-19. Real time RT-PCR is the only laboratory test available till now to confirm the infection. However, the accuracy of real time RT-PCR depends on many factors, including sampling location and of methods, quality of RNA extraction and training of operators etc.. Variations in these factors might significantly lower the sensitivity of the detection. We developed a peptide-based luminescent immunoassay to detect IgG and IgM. Cut-off value of this assay was determined by the detection of 200 healthy sera and 167 sera from patients infected with other pathogens than SARS-CoV-2. To evaluate the performance of this assay, we detected IgG and IgM in the 276 sera from confirmed patients. The positive rate of IgG and IgM were 71.4% (197/276) and 57.2% (158/276) respectively. By combining with real time RT-PCR detection, this assay might help to enhance the accuracy of diagnosis of SARS-CoV-2 infection.


2018 ◽  
Vol 25 (18) ◽  
pp. 2140-2165 ◽  
Author(s):  
Jelena B. Popovic-Djordjevic ◽  
Ivana I. Jevtic ◽  
Tatjana P. Stanojkovic

Background: Diabetes mellitus type 2 (DMT2) is an endocrine disease of global proportions which is currently affecting 1 in 12 adults in the world, with still increasing prevalence. World Health Organization (WHO) declared this worldwide health problem, as an epidemic disease, to be the only non-infectious disease with such categorization. People with DMT2 are at increased risk of various complications and have shorter life expectancy. The main classes of oral antidiabetic drugs accessible today for DMT2 vary in their chemical composition, modes of action, safety profiles and tolerability. Methods: A systematic search of peer-reviewed scientific literature and public databases has been conducted. We included the most recent relevant research papers and data in respect to the focus of the present review. The quality of retrieved papers was assessed using standard tools. Results: The review highlights the chemical structural diversity of the molecules that have the common target-DMT2. So-called traditional antidiabetics as well as the newest and the least explored drugs include polypeptides and amino acid derivatives (insulin, glucagon-like peptide 1, dipeptidyl peptidase-IV inhibitors, amylin), sulfonylurea derivatives, benzylthiazolidine- 2,4-diones (peroxisome proliferator activated receptor-γ agonists/glitazones), condensed guanido core (metformin) and sugar-like molecules (α-glucosidase and sodium/ glucose co-transporter 2 inhibitors). Conclusion: As diabetes becomes a more common disease, interest in new pharmacological targets is on the rise.


2020 ◽  
Author(s):  
Andrea Bartolini ◽  
Margherita Scapaticci ◽  
Marina Bioli ◽  
Tiziana Lazzarotto ◽  
Maria Carla Re ◽  
...  

In March 2020, the World Health Organization (WHO) declared a pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Due to the absence of effective treatment or biomedical prevention, understanding potential post infection immunity has important implications for epidemiologic assessments. For this reason, increasing number of in vitro diagnostic companies are developing serological assays to detect antibodies against SARS-CoV-2, but most of them lack the validation by third parties in relation to their quality, limiting their usefulness. We submitted to serological screening by two different immunochromatographic (IC) rapid testing for detection of IgG and IgM against SARS-CoV-2, 151 asymptomatic or minimally symptomatic healthcare workers previously tested positive for SARS-CoV-2 RT-PCR in order to evaluate the performance of rapid assays. Results showed discrepancies between molecular and IC results, and an inconsistency of immunoglobulins positivity patterns when compared to ELISA/CLIA results, highlighting the absolute necessity of assays performance validation before their marketing and use, in order to avoid errors in the results evaluation at both clinical and epidemiological level.


2021 ◽  
pp. 2979-2983
Author(s):  
Hamong Suharsono ◽  
Ali Ghufron Mukti ◽  
Ketut Suryana ◽  
I. Wayan Masa Tenaya ◽  
Dilasdita Kartika Pradana ◽  
...  

Background and Aim: Coronavirus disease 2019 (COVID-19) is an acute infectious respiratory disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and has spread rapidly globally, resulting in a pandemic. In humans, the main routes of transmission are respiratory droplets and close contact with infected individuals or through contact with an object infected with the virus, followed by touching mouth, nose, or eyes. It is assumed that SARS-CoV-2 was originated in wild animals and was then transmitted to humans. Although some wildlife and domestic animals can be naturally or experimentally infected with the virus, the intermediate hosts that transmitted it to humans are still unknown. Understanding the dynamics of SARS-CoV-2 associated with possible zoonotic transmission of intermediate hosts is considered critical. Reportedly, cats or dogs living with COVID-19-positive humans tested positive for the disease, suggesting that the virus was transmitted to the animals from humans. Information regarding the epidemiological investigation and comprehensive studies is limited. Therefore, it is still unclear how high is the correlation of infection in humans and pet animals, especially those living together. The aim of this study was to investigate the possibility of SARS-CoV-2 infection in the pets of patients with COVID-19 who were hospitalized at the Wangaya hospital, Denpasar, Bali, Indonesia. Materials and Methods: A total of seven clinically asymptomatic pets (six dogs of different races and sexes and a cat [age, 360-2920 days]) were included in this study. These animals belonged to patients with confirmed SARS-CoV-2 infection from August to November 2020. Nasal swab and nasopharyngeal samples were collected from the pets individually under anesthetic condition and were collected 6-12 days after confirmed SARS-CoV-2 infection in owners and hospitalization at the Wangaya Hospital. The swab samples were then processed for RNA isolation and tested using reverse transcription-polymerase chain reaction (RT-PCR) for SARS-CoV-2, in accordance with the World Health Organization manual 2020. Results: RT-PCR results for all seven RNA samples, prepared from the swab samples, were negative. For the samples, all PCR products were below the threshold limit, suggesting no genetic material belonging to the samples tested. Conclusion: This was the first preliminary study of COVID-19 on pets in pandemic using RT-PCR. The study tested a very limited quantity of samples, and all of them were negative. However, the way in which the samples were prepared was considered appropriate. Therefore, in further studies, testing of more samples of pets of more individuals with confirmed SARS-CoV-2 infection is required.


Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1384
Author(s):  
Michael Zapor

The Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV-2) that emerged in December 2019 as the causative agent of Coronavirus 2019 (COVID-19) and was declared a pandemic by the World Health Organization in March 2020 has several distinctive features, including extensive multiorgan involvement with a robust systemic inflammatory response, significant associated morbidity and mortality, and prolonged persistence of viral RNA in the clinical specimens of infected individuals as detected by Reverse Transcription Polymerase Chain Reaction (RT-PCR) amplification. This review begins with an overview of SARS-CoV-2 morphology and replication and summarizes what is known to date about the detection of the virus in nasal, oropharyngeal, and fecal specimens of patients who have recovered from COVID-19, with a focus on the factors thought to contribute to prolonged detection. This review also provides a discussion on the infective potential of this material from asymptomatic, pre-symptomatic, and convalescing individuals, to include a discussion of the relative persistence and infectious potential of virus in clinical specimens recovered from pediatric COVID-19 patients.


2021 ◽  
Vol 2 (4) ◽  
pp. 431-441
Author(s):  
Caio Ricardo Eich ◽  
Barbara Scariot Colombelli ◽  
Kattlyn Larissa Candido ◽  
Luciana Oliveira De Fariña

Em 11 de março de 2020, a Organização Mundial de Saúde (OMS) decretou a pandemia do COVID-19, causado pelo vírus SARS-CoV-2, responsável por mais de 4,5 milhões de mortes até o momento. Esta nova realidade exigiu respostas por parte das autoridades e da população, a fim de mitigar a propagação do vírus e evitar o colapso do sistema de saúde, assim como estudos de vigilância em saúde, que possibilitaram um melhor entendimento dos mecanismos de transmissão do vírus e possibilitaram identificar zonas de risco dentro de cidades ou ambientes públicos. Este estudo tem o objetivo de identificar a presença do SARS-CoV-2 dentro da Universidade Estadual do Oeste do Paraná, a qual fornece serviços de saúde para a população local, assim como realizar um controle interno no Laboratório de Bioquímica Molecular (LaBioqMol) da universidade, onde são realizados testes de RT-PCR semanalmente. Foram coletadas 21 amostras de áreas frequentemente tocadas por pessoas, cuja presença do RNA viral e de material genético humano foi identificada por RT-PCR. Em nenhuma das amostras foi detectado a presença do vírus. Entretanto, em 8 (38,1%) das amostras foi verificada a amplificação do gene RNaseP, indicando a presença de células humana. Este estudo auxilia no controle e garantia de qualidade do LaBioqMol e fortalece a visão de que a contaminação do ambiente pelo SARS-CoV-2 é provavelmente menos frequente do que foi anteriormente sugerido no início da pandemia.   On March 11, 2020, the World Health Organization (WHO) decreed the pandemic of COVID-19, caused by the SARS-CoV-2 virus, responsible for more than 4.5 million deaths to date. This new reality demanded responses from the authorities and the population in order to mitigate the spread of the virus and avoid the collapse of the health system, as well as health surveillance studies, which enabled a better understanding of the mechanisms of virus transmission and made it possible to identify risk zones within cities or public environments. This study aims to identify the presence of SARS-CoV-2 within the Universidade Estadual do Oeste do Paraná, which provides health services to the local population, as well as to perform an internal control at the university's Molecular Biochemistry Laboratory (LaBioqMol), where RT-PCR tests are performed weekly. Twenty-one samples were collected from areas frequently touched by people, and the presence of viral RNA and human genetic material was identified by RT-PCR. In none of the samples was the presence of the virus detected. However, in 8 (38.1%) of the samples the RNaseP gene amplification was verified, indicating the presence of human cells. This study assists in quality control and assurance at LaBioqMol and strengthens the view that environmental contamination by SARS-CoV-2 is probably less frequent than was previously suggested at the beginning of the pandemic.


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