scholarly journals Influence of viral transport media and freeze-thaw cycling on the sensitivity of qRT-PCR detection of SARS-CoV-2 nucleic acids

Nanoscale ◽  
2021 ◽  
Author(s):  
Cian Holohan ◽  
Sophia Hanrahan ◽  
Nathan Feely ◽  
Peng Li ◽  
John O'Connell ◽  
...  
Keyword(s):  
Infectious Diseases ◽  
Nucleic Acids ◽  
Large Scale ◽  
Diagnostic Testing ◽  
Pcr Detection ◽  
Molecular Diagnostic ◽  
Freeze Thaw ◽  
Qrt Pcr ◽  
Viral Transport ◽  
Molecular Diagnostic Testing

Objective The events of the last year have highlighted the complexity of implementing molecular diagnostic testing for infectious diseases at a large scale. The purpose of this study was to...

scholarly journals Influence of viral transport media and freeze-thaw cycling on the sensitivity of qRT-PCR detection of SARS-CoV-2 nucleic acids

2021 ◽  
Author(s):  
Cian Holohan ◽  
Sophia Hanrahan ◽  
Nathan Feely ◽  
Peng Li ◽  
John O'Connell ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Large Scale ◽  
Pcr Detection ◽  
Molecular Diagnostic ◽  
Pcr Analysis ◽  
Analytical Sensitivity ◽  
Freeze Thaw ◽  
Qrt Pcr ◽  
Viral Transport ◽  
The Stability

The events of the last year have highlighted the complexity of implementing large-scale molecular diagnostic testing for novel pathogens. The purpose of this study was to determine the chemical influences of sample collection media and storage on the stability and detection of viral nucleic acids by qRT-PCR. We studied the mechanism(s) through which viral transport media (VTM) and number of freeze-thaw cycles influenced the analytical sensitivity of qRT-PCR detection of SARS-CoV-2. Our goal is to reinforce testing capabilities and identify weaknesses that could arise in resource-limited environments that do not have well-controlled cold chains. The sensitivity of qRT-PCR analysis was studied in four VTM for synthetic single-stranded RNA (ssRNA) and double-stranded DNA (dsDNA) simulants of the SARS-CoV-2 genome. The sensitivity and reproducibility of qRT-PCR for the synthetic ssRNA and dsDNA were found to be highly sensitive to VTM with the best results observed for ssRNA in HBSS and PBS-G. Surprisingly, the presence of epithelial cellular material with the ssRNA increased the sensitivity of the qRT-PCR assay. Repeated freeze-thaw cycling decreased the sensitivity of the qRT-PCR with two noted exceptions. The choice of VTM is critically important to defining the sensitivity of COVID-19 molecular diagnostics assays and this study suggests they can impact upon the stability of the SARS-CoV-2 viral genome. This becomes increasingly important if the virus structure is destabilised before analysis, which can occur due to poor storage conditions. This study suggests that COVID-19 testing performed with glycerol-containing PBS will produce a high level of stability and sensitivity. These results are in agreement with clinical studies reported for patient-derived samples.

Analysis of the tendency of development of biosensors based on GaN

2021 ◽  
Author(s):  
S.I. Agasieva ◽  
E.A. Smetanin ◽  
A.R. Vechkanov ◽  
A.V. Gubanov
Keyword(s):  
Clinical Trials ◽  
Infectious Diseases ◽  
Diagnostic Tests ◽  
Diagnostic Testing ◽  
Healthcare Systems ◽  
Clinical Analysis ◽  
Molecular Diagnostic ◽  
Economic Damage ◽  
Disease Research ◽  
Local Healthcare

Statement of the problem of this article - one of the most important problems is protection from especially dangerous infectious diseases. The use of biosensors in clinical trials will significantly reduce the time for obtaining the results of analyzes, thereby speeding up the appointment of treatment to patients. The purpose of the article is to present modern designs of biosensors based on gallium nitride, the possibilities of their application and characteristics. Consider the principles of operation, areas of application and characteristics. As a result, the design of modern biosensors and modern trends in their use from various sources of literature in recent years are shown. Biosensors, principles of their action, areas of application and characteristics are considered, which will reduce the possible socio-economic damage from temporary disability for sick citizens due to the rapid and timely implementation of anti-epidemic measures. Practical value: the proposed biosensors are of interest as devices for detecting diseases. The use of biosensors in clinical disease research has several potential advantages over other clinical analysis methods, including increased analysis speed and flexibility, multipurpose analysis capability, automation, reduced diagnostic testing costs, and the ability to integrate molecular diagnostic tests into local healthcare systems.

scholarly journals Large-scale, in-house production of viral transport media to support SARS-CoV-2 PCR testing in a multi-hospital healthcare network during the COVID-19 pandemic

2020 ◽  
Author(s):  
Kenneth P Smith ◽  
Annie Cheng ◽  
Amber Chopelas ◽  
Sarah DuBois-Coyne ◽  
Ikram Mezghani ◽  
...  
Keyword(s):  
Large Scale ◽  
Medical Center ◽  
Diagnostic Testing ◽  
Diagnostic Laboratory ◽  
Healthcare Network ◽  
Research Division ◽  
Viral Transport ◽  
Accelerated Stability ◽  
House Production ◽  
Transport Tube

The COVID-19 pandemic has severely disrupted worldwide supplies of viral transport media (VTM) due to widespread demand for SARS-CoV-2 RT-PCR testing. In response to this ongoing shortage, we began production of VTM in-house in support of diagnostic testing in our hospital network. As our diagnostic laboratory was not equipped for reagent production, we took advantage of space and personnel that became available due to closure of the research division of our medical center. We utilized a formulation of VTM described by the CDC that was simple to produce, did not require filtration for sterilization, and used reagents that were available from commercial suppliers. Performance of VTM was evaluated by several quality assurance measures. Based on Ct values of spiking experiments, we found that our VTM supported highly consistent amplification of the SARS-CoV-2 target (coefficient of variation = 2.95%) using the Abbott RealTime SARS-CoV-2 EUA assay on the Abbott m2000 platform. VTM was also found to be compatible with multiple swab types and, based on accelerated stability studies, able to maintain functionality for at least four months at room temperature. We further discuss how we met logistical challenges associated with large-scale VTM production in a crisis setting including use of staged, assembly line for VTM transport tube production.

scholarly journals A Report on Molecular Diagnostic Testing for Inherited Retinal Dystrophies by Targeted Genetic Analyses

2017 ◽  
Vol 21 (2) ◽  
pp. 66-73
Author(s):  
Hema L. Ramkumar ◽  
Harini V. Gudiseva ◽  
Kameron T. Kishaba ◽  
John J. Suk ◽  
Rohan Verma ◽  
...  
Keyword(s):  
Diagnostic Testing ◽  
Molecular Diagnostic ◽  
Genetic Analyses ◽  
Inherited Retinal Dystrophies ◽  
Retinal Dystrophies ◽  
Molecular Diagnostic Testing
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