scholarly journals Reliable Diagnostics of SARS-CoV-2 Infections Using One- and Two-Gene Molecular Tests for a Viral RNA Detection—Results Questioning Previous Observations

Diagnostics ◽  
2021 ◽  
Vol 11 (10) ◽  
pp. 1839
Author(s):  
Tomasz Bogiel ◽  
Mateusz Rzepka ◽  
Dagmara Depka
Keyword(s):  
Target Genes ◽  
Medical Problem ◽  
Viral Rna ◽  
Becton Dickinson ◽  
Rna Detection ◽  
Molecular Tests ◽  
Negative Results ◽  
Rapid Spread ◽  
Novel Coronavirus ◽  
Biotechnology Products

SARS-CoV-2 is a new virus from the Coronaviridae family and its rapid spread is now the most important medical problem worldwide. Currently used tests vary in the number and selection of SARS-CoV-2 target genes. Meanwhile, the choice of the appropriate target gene may be important in terms of a reliable detection of a viral RNA. As some researchers questioned the sensitivity of the monogenic VIASURE SARS-CoV-2 S gene Real Time PCR Detection Kit (CerTest Biotec, Zaragoza, Spain) in mid-2020, the aim of the study was to evaluate the usefulness of this kit, used along with the BD MAX™ System (Becton Dickinson, East Rutherford, NJ, USA), and compare the results with two-gene Bosphore Novel Coronavirus (2019-nCoV) Detection Kit v1 (Anatolia Diagnostics and Biotechnology Products Inc., Istanbul, Turkey). Both tests were carried out on 306 nasopharyngeal/oropharyngeal swabs. The consistent results (72 positive and 225 negative results found simultaneously in both kits) were obtained for 297 (97.1%) samples altogether, while discrepancies between the results of the evaluated tests were observed for nine (2.9%) specimens. There were no statistically significant differences between the method used and the frequency of positive results. Both tests, targeted at detecting one and two genes, are effective in SARS-CoV-2 RNA detection.

scholarly journals Superiority of MALDI-TOF Mass Spectrometry over Real-Time PCR for SARS-CoV-2 RNA Detection

Viruses ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 730
Author(s):  
Magda Rybicka ◽  
Ewa Miłosz ◽  
Krzysztof Piotr Bielawski
Keyword(s):  
Mass Spectrometry ◽  
Gold Standard ◽  
Viral Genome ◽  
False Negative ◽  
Rt Pcr ◽  
Rna Detection ◽  
Maldi Tof ◽  
Negative Results ◽  
False Negative Results ◽  
Pcr Test

At present, the RT-PCR test remains the gold standard for early diagnosis of SARS-CoV-2. Nevertheless, there is growing evidence demonstrating that this technique may generate false-negative results. Here, we aimed to compare the new mass spectrometry-based assay MassARRAY® SARS-CoV-2 Panel with the RT-PCR diagnostic test approved for clinical use. The study group consisted of 168 suspected patients with symptoms of a respiratory infection. After simultaneous analysis by RT-PCR and mass spectrometry methods, we obtained discordant results for 17 samples (10.12%). Within fifteen samples officially reported as presumptive positive, 13 were positive according to the MS-based assay. Moreover, four samples reported by the officially approved RT-PCR as negative were positive in at least one MS assay. We have successfully demonstrated superior sensitivity of the MS-based assay in SARS-CoV-2 detection, showing that MALDI-TOF MS seems to be ideal for the detection as well as discrimination of mutations within the viral genome.

scholarly journals A new model for COVID-19 control and its implementation in the city of Honghu, China: a case report

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
He-ran Wang ◽  
Meng-chun Gong ◽  
Jing-Yuan Sun ◽  
Jian Sun ◽  
Yi Guo ◽  
...  
Keyword(s):  
Management Strategy ◽  
Human Infection ◽  
Public Health Emergency ◽  
Medical Resources ◽  
Case Presentation ◽  
Management Measures ◽  
Rapid Spread ◽  
Medical Teams ◽  
Novel Coronavirus ◽  
The City

Abstract Background Novel coronavirus pneumonia has been the most serious worldwide public health emergency since being identified in December 2019. The rapid spread of the pandemic and the strong human to human infection rate of COVID-19 poses a great prevention challenge. There has been an explosion in the number of confirmed cases in several cities near Wuhan, including the highest in Honghu, Jinzhou. Owing to the limited admission capacity and medical resources, increasing numbers of suspected cases of COVID-19 infection were difficult to confirm or treat. Case presentation Following the arrival of the Guangdong medical aid team on 11 February, 2020, COVID-19 care in Honghu saw changes after a series of solutions were implemented based on the ‘Four-Early’ and ‘Four-centralization’ management measures. The ‘Four-Early’ measures are: early detection, early reporting, early quarantine, and early treatment for meeting an urgent need like the COVID-19 pandemic. ‘Four-centralization’ refers to the way in which recruited medical teams can make full use of medical resources to give patients the best treatment. These solutions successfully increased the recovery rate and reduced mortality among patients with COVID-19 in Honghu. Conclusions This management strategy is called the ‘Honghu Model’ which can be generalized to enable the prevention and management of COVID-19 worldwide.

scholarly journals Do I Have HIV or Not? Lack of RNA Detection and the Case for Sensitive DNA Testing

2020 ◽  
Vol 7 (11) ◽  
Author(s):  
Sandra A Springer ◽  
Silvina Masciotra ◽  
Jeffrey A Johnson ◽  
Sheldon Campbell
Keyword(s):  
Antiretroviral Therapy ◽  
Hiv Infection ◽  
Viral Rna ◽  
Infection Status ◽  
Test Results ◽  
Dna Testing ◽  
Rna Detection ◽  
Hiv Test

Abstract We present a case of a 20-year-old male who had ambiguous HIV test results after entering new provider care and whose status was later complicated by undetectable viral RNA off antiretroviral therapy (ART). Verifying HIV infection status may occasionally require sensitive DNA testing that might need to be considered in diagnostic guidelines to resolve diagnosis and ensure appropriate ART management.

scholarly journals Previremic Identification of Ebola or Marburg Virus Infection Using Integrated Host-Transcriptome and Viral Genome Detection

mBio ◽  
2020 ◽  
Vol 11 (3) ◽  
Author(s):  
Emily Speranza ◽  
Ignacio Caballero ◽  
Anna N. Honko ◽  
Joshua C. Johnson ◽  
J. Kyle Bohannon ◽  
...  
Keyword(s):  
Host Response ◽  
Nonhuman Primates ◽  
Ebola Virus ◽  
Clinical Symptoms ◽  
Infectious Agent ◽  
Marburg Virus ◽  
Viral Rna ◽  
Molecular Tests ◽  
Early Replication ◽  
Host Infection

ABSTRACT Outbreaks of filoviruses, such as those caused by the Ebola (EBOV) and Marburg (MARV) virus, are difficult to detect and control. The initial clinical symptoms of these diseases are nonspecific and can mimic other endemic pathogens. This makes confident diagnosis based on clinical symptoms alone impossible. Molecular diagnostics for these diseases that rely on the detection of viral RNA in the blood are only effective after significant disease progression. As an approach to identify these infections earlier in the disease course, we tested the effectiveness of viral RNA detection combined with an assessment of sentinel host mRNAs that are upregulated following filovirus infection. RNAseq analysis of EBOV-infected nonhuman primates identified host RNAs that are upregulated at early stages of infection. NanoString probes that recognized these host-response RNAs were combined with probes that recognized viral RNA and were used to classify viral infection both prior to viremia and postviremia. This approach was highly successful at identifying samples from nonhuman primate subjects and correctly distinguished the causative agent in a previremic stage in 10 EBOV and 5 MARV samples. This work suggests that unified host response/viral fingerprint assays can enable diagnosis of disease earlier than testing for viral nucleic acid alone, which could decrease transmission events and increase therapeutic effectiveness. IMPORTANCE Current molecular tests that identify infection with high-consequence viruses such as Ebola virus and Marburg virus are based on the detection of virus material in the blood. These viruses do not undergo significant early replication in the blood and, instead, replicate in organs such as the liver and spleen. Thus, virus begins to accumulate in the blood only after significant replication has already occurred in those organs, making viremia an indicator of infection only after initial stages have become established. Here, we show that a multianalyte assay can correctly identify the infectious agent in nonhuman primates (NHPs) prior to viremia through tracking host infection response transcripts. This illustrates that a single-tube, sample-to-answer format assay could be used to advance the time at which the type of infection can be determined and thereby improve outcomes.

scholarly journals Wellness Constraints Among International University Students Transitioning Through Covid 19

2021 ◽  
Vol 58 (3) ◽  
pp. 3444-3456
Author(s):  
Mr J Dorasamy, Et. al.
Keyword(s):  
World Health Organization ◽  
Social Economic ◽  
World Health ◽  
Social Emotional ◽  
Global Pandemic ◽  
Rapid Spread ◽  
The World ◽  
International University ◽  
Novel Coronavirus ◽  
Health Organization

The World Health Organization (Who) In March 2020 Declared Covid 19 A Pandemic, Due To The  Global And Rapid Spread Of A Novel Coronavirus (Who, 2020). The Covid 19 Pandemic Being Highly Infectious And Unpredictable, Has  Disrupted  Social, Economic, Environmental And Political Spheres Of Life. Globally, People Have Ventured Into A “Lockdown World”, Increasing Uncertainty About Their Future Amidst The Covid 19 Pandemic. As A Result Of The Pandemic, Social Alteration Has Taken The Form Of Social Distancing, Self-Isolation And Self-Quarantine.  Many Were Unprepared For The Shift From The “Normal”, Propelling  Undue  Stress Under The New Normal Way Of Doing Things During The Current Global Pandemic Crisis. This Has Been Accompanied By Social, Emotional And Mental Effects, As The Ongoing And Fluid Nature Of The Pandemic Has Created Uncertainty For Many People. The Covid 19 Pandemic, As A Multidimensional Stressor Affecting Wellbeing, Has Affected Individuals, Families, Educational, Occupational, And Broader Societal Systems.  

scholarly journals Understanding the Use of Real Time Reverse Transcriptase-Polymerase Chain Reaction (rRT-PCR) For Covid-19 Diagnosis

2020 ◽  
Vol 4 (2) ◽  
pp. 86-97
Author(s):  
Adesola O. Olalekan ◽  
Bamidele A. Iwalokun ◽  
Olutoyin C. Adekunle ◽  
Hussaini A. Makun ◽  
Tatfeng Mirabeau ◽  
...  
Keyword(s):  
Reverse Transcriptase ◽  
Real Time ◽  
Target Genes ◽  
Protein Gene ◽  
Rna Detection ◽  
Glycoprotein Gene ◽  
Adequate Knowledge ◽  
Alternative Testing ◽  
Polymerase Chain ◽  
Testing Protocols

Background: Adequate knowledge of real time Reverse Transcriptase-Polymerase Chain Re-action (rRT-PCR) is critical for accurate implementation of the assay, interpretation of results and report-ing. This mini-review describes the principles, procedures, and level of development of rRT-PCR assays for the control of the COVID-19 pandemic. Methods: A narrative review was carried out to describe the principles of rRT-PCR, provide an update on the landscape of rRT-PCR protocols and elucidate the process control involved in pre-analytical, analytical and post-analytical stages of COVID-19 testing . Review Findings: The rRT-PCR is currently considered to be the acceptable standard for confirming COVID-19 diagnosis based on SARS-CoV-2 RNA detection via conversion to cDNA and amplification of target genes in real time using sequence specific TaqMan® probes. Available evidence indicates that different rRT-PCR protocols varying in number and type of target genes within SARS-CoV-2 genome are currently available for validation and emergency use approval (EUA) in pandemic countries. A total of 1 – 3 target genes, comprising the ORF1a, ORF1b, RNA dependent RNA polymerase (RdRp), Nucleoplasid protein gene (N), Spike glycoprotein gene (S) and Envelope protein gene (E) are detected by these proto-cols. Conclusion: rRT-PCR remains the most sensitive method for confirming, monitoring and managing COVID-19 disease in the ongoing pandemic in all affected countries. The need for validation of every rRT-PCR protocol prior to deployment for COVID-19 testing and research into the development of alternative testing protocols are strongly recommended

scholarly journals 2019 Novel Coronavirus (COVID-19) Outbreak: A Review of the Current Literature and Built Environment (BE) Considerations to Reduce Transmission

2020 ◽  
Author(s):  
Leslie Dietz ◽  
Patrick F. Horve ◽  
David Coil ◽  
Mark Fretz ◽  
Jonathan Eisen ◽  
...  
Keyword(s):  
Built Environment ◽  
Assisted Living ◽  
Disease Transmission ◽  
Health Care Facilities ◽  
Federal State ◽  
Public Administrators ◽  
Infectious Disease Transmission ◽  
City Governments ◽  
Rapid Spread ◽  
Novel Coronavirus

With the rapid spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that results in coronavirus disease 2019 (COVID-19), corporate entities, federal, state, county and city governments, universities, school districts, places of worship, prisons, health care facilities, assisted living organizations, daycares, homeowners, and other building owners and occupants have an opportunity to reduce the potential for transmission through built environment (BE) mediated pathways. Over the last decade, substantial research into the presence, abundance, diversity, function, and transmission of microbes in the BE has taken place and revealed common pathogen exchange pathways and mechanisms. In this paper, we synthesize this microbiology of the BE research and the known information about SARS-CoV-2 to provide actionable and achievable guidance to BE decision makers, building operators, and all indoor occupants attempting to minimize infectious disease transmission through environmentally mediated pathways. We believe this information is useful to corporate and public administrators and individuals responsible for building operations and environmental services in their decision-making process about the degree and duration of social-distancing measures during viral epidemics and pandemics.

scholarly journals Oral aspects related to SARS-CoV-2

2021 ◽  
Vol 1 (2) ◽  
pp. 153-160
Author(s):  
Akram Belmehdi ◽  
Saliha Chbicheb
Keyword(s):  
Oral Cavity ◽  
Diagnostic Value ◽  
Disease Spread ◽  
Health Crisis ◽  
Rapid Spread ◽  
The World ◽  
Spanish Flu ◽  
Taste Disorders ◽  
Novel Coronavirus ◽  
The Relationship

The pandemic of coronavirus disease (COVID-19) is considered as the biggest global health crisis for the world since the Spanish flu, also known as the 1918 flu pandemic. Driven by the SARS-CoV-2 novel coronavirus infection, the rapid spread of this disease and the related pneumonia COVID-19 are a challenge for healthcare systems in over the world, and it is a constantly evolving situation with new symptoms and prognostic factors. SARS-CoV-2 has lately been detected in infected patient’s oral cavity; the COVID-19 outbreak is an alert that all dental and other health professionals must be vigilant in defending against the infectious disease spread. This review summarizes an update from current medical literature about the relationship between oral cavity and coronavirus disease by presenting some oral aspects which was detected in infected patients such as the oral lesions related to this virus and its therapeutic protocol, taste disorders and also the diagnostic value of saliva for SARS-CoV-2.

scholarly journals Direct Viral RNA Detection of SARS-CoV-2 and DENV in Inactivated Samples by Real-Time RT-qPCR: Implications for Diagnosis in Resource Limited Settings with Flavivirus Co-Circulation

Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1558
Author(s):  
Zhan Qiu Mao ◽  
Mizuki Fukuta ◽  
Jean Claude Balingit ◽  
Thi Thanh Ngan Nguyen ◽  
Co Thach Nguyen ◽  
...  
Keyword(s):  
Real Time ◽  
Early Stage ◽  
Rna Extraction ◽  
Viral Rna ◽  
Clinical Samples ◽  
Heat Inactivation ◽  
Resource Limited Settings ◽  
Rna Detection ◽  
Resource Limited ◽  
Starting Point

The RT-qPCR method remains the gold standard and first-line diagnostic method for the detection of SARS-CoV-2 and flaviviruses, especially in the early stage of viral infection. Rapid and accurate viral detection is a starting point in the containment of the COVID-19 pandemic and flavivirus outbreaks. However, the shortage of diagnostic reagents and supplies, especially in resource-limited countries that experience co-circulation of SARS-CoV-2 and flaviviruses, are limitations that may result in lesser availability of RT-qPCR-based diagnostic tests. In this study, the utility of RNA-free extraction methods was assessed for the direct detection of SARS-CoV-2 and DENV-2 in heat-inactivated or chemical-inactivated samples. The findings demonstrate that direct real-time RT-qPCR is a feasible option in comparison to conventional real-time RT-qPCR based on viral genome extraction-based methods. The utility of heat-inactivation and direct real-time RT-qPCR for SARS-CoV-2, DENV-2 viral RNA detection was demonstrated by using clinical samples of SARS-CoV-2 and DENV-2 and spiked cell culture samples of SARS-CoV-2 and DENV-2. This study provides a simple alternative workflow for flavivirus and SARS-CoV-2 detection that includes heat inactivation and viral RNA extraction-free protocols, with aims to reduce the risk of exposure during processing of SARS-CoV-2 biological specimens and to overcome the supply-chain bottleneck, particularly in resource limited settings with flavivirus co-circulation.

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