scholarly journals The Production of Standardized Samples with Known Concentrations for Severe Acute Respiratory Syndrome Coronavirus 2 RT-qPCR Testing Validation for Developing Countries in the Period of the Pandemic Era

2021 ◽  
Vol 2021 ◽  
pp. 1-10
Author(s):  
Hoang Quoc Cuong ◽  
Nguyen Duc Hai ◽  
Hoang Thuy Linh ◽  
Nguyen Trung Hieu ◽  
Nguyen Hoang Anh ◽  
...  

Background. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a pandemic of pneumonia spreading around the world, leading to serious threats to public health and attracting enormous attention. There is an urgent need for sensitive diagnostic testing implementation to control and manage SARS-CoV-2 in public health laboratories. The quantitative reverse transcription PCR (RT-qPCR) assay is the gold standard method, but the sensitivity and specificity of SARS-CoV-2 testing are dependent on a number of factors. Methods. We synthesized RNA based on the genes published to estimate the concentration of inactivated virus samples in a biosafety level 3 laboratory. The limit of detection (LOD), linearity, accuracy, and precision were evaluated according to the bioanalytical method validation guidelines. Results. We found that the LOD reached around 3 copies/reaction. Furthermore, intra-assay precision, accuracy, and linearity met the accepted criterion with an RSD for copies of less than 25%, and linear regression met the accepted R 2 of 0.98. Conclusions. We suggest that synthesized RNA based on the database of the NCBI gene bank for estimating the concentration of inactivated virus samples provides a potential opportunity for reliable testing to diagnose coronavirus disease 2019 (COVID-19) as well as limit the spread of the disease. This method may be relatively quick and inexpensive, and it may be useful for developing countries during the pandemic era. In the long term, it is also applicable for evaluation, verification, validation, and external quality assessment.

2000 ◽  
Vol 66 (10) ◽  
pp. 4383-4388 ◽  
Author(s):  
P. W. Huang ◽  
D. Laborde ◽  
V. R. Land ◽  
D. O. Matson ◽  
A. W. Smith ◽  
...  

ABSTRACT Human caliciviruses (HuCVs) cause waterborne outbreaks of gastroenteritis. Standard indicators of a safe water supply do not adequately predict contamination of water by viruses, including HuCVs. We developed a method to concentrate and detect HuCVs in water samples by using a cultivable primate calicivirus (Pan-1) as a model. Viable Pan-1 was seeded in different types of water and then filtered with a 1MDS filter, eluted with beef extract (BE), and reconcentrated by polyethylene glycol (PEG) precipitation. The viruses in the final samples were tested by plaque assay or by reverse transcription (RT)-PCR following extraction of the RNA with Trizol. Pan-1 was more sensitive to high-pH treatment than poliovirus was; a pH 9.0 BE solution was found to recover 35% more viable Pan-1 than a pH 9.5 BE solution recovered. Pan-1 was recovered from small volumes of deionized, finished, ground, and surface waters at efficiencies of 94, 73, 67, and 64%, respectively, when samples were assayed after elution without further concentration. When larger volumes of water (up to 40 liters) were tested after elution and concentration with PEG, 38, 19, and 14% of the seeded Pan-1 were recovered from finished, ground, and surface waters, respectively. The limit of detection of Pan-1 by RT-PCR was estimated to be 0.75 to 1.5 PFU in 40 liters of finished water. This method may be adapted for monitoring HuCVs in drinking water and other types of water for public health safety.


2021 ◽  
Author(s):  
Tran Bac Le ◽  
Hye Kwon Kim ◽  
Min-Ju Ahn ◽  
Mark Zanin ◽  
Van Thi Lo ◽  
...  

Abstract The pandemic coronavirus disease 2019 (COVID-19) is characterized as an acute respiratory infection in the majority of cases and is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). However, other coronaviruses (CoVs) can infect humans, although the majority only cause mild respiratory symptoms. As early diagnosis of SARS-CoV-2 is critical to prevent further transmission events and to improve clinical outcomes, it is important to be able to distinguish SARS-CoV-2 from other CoVs in respiratory samples. Therefore, we developed and evaluated a novel quantitative reverse transcription PCR (RT-qPCR) assay, targeting the spike (S) and membrane (M) genes, to enable the rapid identification of SARS-CoV-2 including several new circulating variants, and other emerging pan-SARS-like CoVs. Using RNA extracted from cell culture supernatants, our multiple simultaneous SARS-CoV-2 assays confirmed a limit of detection of 1 × 100 TCID50/ml and no cross-reaction with human coronaviruses or other respiratory viruses. We also validated our method using human clinical samples from COVID-19 patients and healthy individuals, including nasal swab and sputum samples. This novel one-step multiplex RT-qPCR assay can be used to improve the laboratory diagnosis of human-pathogenic coronaviruses, including SARS-CoV-2, and may be useful for the identification of other pan-SARS-like CoVs of zoonotic origin.


Author(s):  
Kunal Parikh ◽  
Tanvi Makadia ◽  
Harshil Patel

Dengue is unquestionably one of the biggest health concerns in India and for many other developing countries. Unfortunately, many people have lost their lives because of it. Every year, approximately 390 million dengue infections occur around the world among which 500,000 people are seriously infected and 25,000 people have died annually. Many factors could cause dengue such as temperature, humidity, precipitation, inadequate public health, and many others. In this paper, we are proposing a method to perform predictive analytics on dengue’s dataset using KNN: a machine-learning algorithm. This analysis would help in the prediction of future cases and we could save the lives of many.


2017 ◽  
Vol 25 (1) ◽  
pp. 47-65
Author(s):  
Tapiwa V. Warikandwa ◽  
Patrick C. Osode

The incorporation of a trade-labour (standards) linkage into the multilateral trade regime of the World Trade Organisation (WTO) has been persistently opposed by developing countries, including those in Africa, on the grounds that it has the potential to weaken their competitive advantage. For that reason, low levels of compliance with core labour standards have been viewed as acceptable by African countries. However, with the impact of WTO agreements growing increasingly broader and deeper for the weaker and vulnerable economies of developing countries, the jurisprudence developed by the WTO Panels and Appellate Body regarding a trade-environment/public health linkage has the potential to address the concerns of developing countries regarding the potential negative effects of a trade-labour linkage. This article argues that the pertinent WTO Panel and Appellate Body decisions could advance the prospects of establishing a linkage of global trade participation to labour standards without any harm befalling developing countries.


1995 ◽  
Vol 32 (9-10) ◽  
pp. 257-266 ◽  
Author(s):  
G. Kocasoy

Among the different impacts of coastal tourism on the environment, its effect on public health has a special significance. The large increase in population during the high season usually results to an increase in the amount of wastewater produced. If the necessary infrastructure - which most of the time the developing countries lack - for handling these wastes is not available, the sea is polluted causing serious problems. To investigate the effects of sea pollution onpublic health a survey has been conducted in Bodrum and Çeşme; the two most important touristic resorts in Turkey. A questionnaire about the health problems which tourists suffer as well as their culinary habits, swimming habits etc. was distributed. At the same time the pollution level in the sea was determined in the form of coliform concentration. As a result, it was proved that those swimming in polluted waters suffer from gastro-intestinal diseases more than others. Furthermore foreigners and children had been proved to be more susceptible to pollution due to the lack of immunity. The purpose of the present study is to investigate the effect of the rate of pollution as well as of the microbial self purification capacity (T90) on public health.


Author(s):  
Tatsuya Yoshihara ◽  
Kazuya Ito ◽  
Masayoshi Zaitsu ◽  
Eunhee Chung ◽  
Izumi Aoyagi ◽  
...  

Coronavirus disease 2019 (COVID-19) has become a serious public health problem worldwide. In general, healthcare workers are considered to be at higher risk of COVID-19 infection. However, the prevalence of COVID-19 among healthcare workers in Japan is not well characterized. In this study, we aimed to examine the seroprevalence of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) antibodies among 2160 healthcare workers in hospitals and clinics that are not designated to treat COVID-19 patients in Japan. The prevalence of SARS-CoV-2 immunoglobulin G was 1.2% in August and October 2020 (during and after the second wave of the pandemic in Japan), which is relatively higher than that in the general population in Japan (0.03–0.91%). Because of the higher risk of COVID-19 infection, healthcare workers should be the top priority for further social support and vaccination against SARS-CoV-2.


2021 ◽  
Vol 27 (Supplement_1) ◽  
pp. S57-S57
Author(s):  
Edgar Ong ◽  
Ruo Huang ◽  
Richard Kirkland ◽  
Michael Hale ◽  
Larry Mimms

Abstract Introduction A fast (<5 min), time-resolved fluorescence resonance energy transfer (FRET)-based immunoassay was developed for the quantitative detection of infliximab (IFX) and biosimilars for use in therapeutic drug monitoring using only 20 µL of fingerstick whole blood or serum at the point-of-care. The Procise IFX assay and ProciseDx analyzer are CE-marked. Studies were performed to characterize analytical performance of the Procise IFX assay on the ProciseDx analyzer. Methods Analytical testing was performed by spiking known amounts of IFX into negative serum and whole blood specimens. Analytical sensitivity was determined using limiting concentrations of IFX. Linearity was determined by testing IFX across the assay range. Hook effect was assessed at IFX concentrations beyond levels expected to be found within a patient. Testing of assay precision, cross-reactivity and potential interfering substances, and biosimilars was performed. The Procise IFX assay was also compared head-to-head with another CE-marked assay: LISA-TRACKER infliximab ELISA test (Theradiag, France). The accuracy of the Procise IFX assay is established through calibrators and controls traceable to the WHO 1st International Standard for Infliximab (NIBSC code: 16/170). Results The Procise IFX assay shows a Limit of Blank, Limit of Detection, and Lower Limit of Quantitation (LLoQ) of 0.1, 0.2, and 1.1 µg/mL in serum and 0.6, 1.1, and 1.7 µg/mL in whole blood, respectively. The linear assay range was determined to be 1.7 to 77.2 µg/mL in serum and whole blood. No hook effect was observed at an IFX concentration of 200 µg/mL as the value reported as “>ULoQ”. Assay precision testing across 20 days with multiple runs and reagent lots showed an intra-assay coefficient of variation (CV) of 2.7%, an inter-assay CV of <2%, and a total CV of 3.4%. The presence of potentially interfering/cross-reacting substances showed minimal impact on assay specificity with %bias within ±8% of control. Testing of biosimilars (infliximab-dyyb and infliximab-abda) showed good recovery. A good correlation to the Theradiag infliximab ELISA was obtained for both serum (slope=1.01; r=0.99) and whole blood (slope=1.01; r=0.98) samples (Figure 1). Conclusion Results indicate that the Procise IFX assay is sensitive, specific, and precise yielding results within 5 minutes from both whole blood and serum without the operator needing to specify sample type. Additionally, it shows very good correlation to a comparator assay that takes several hours and sample manipulation to yield results. This makes the Procise IFX assay ideal for obtaining fast and accurate IFX quantitation, thus allowing for immediate drug level dosing decisions to be made by the physician during patient treatment.


Author(s):  
Ranjeet S. Sawant ◽  
Bharat D. Zinjurke ◽  
Sandeep V. Binorkar

Abstract The ongoing coronavirus pandemic is caused by severe acute respiratory syndrome coronavirus 2 (SARS CoV 2) and unique in various facets. The earlier experience from the past severe acute respiratory syndrome (SARS) epidemics seem to be insufficient and there is need for better strategies in public health and medical care. Ayurved & Yog are well known for their preventive and therapeutic aspect, but not getting utilized properly for prevention of Covid 19 crisis which may also be helpful as supportive therapy along with current line of management. This paper is aimed at unrevealing the role of Ayurved and Yoga guidelines established by Department of AYUSH for prevention from SARS-CoV-2 by providing help to improving the quality of supportive/prophylactic therapy in relation with their immunity.


Author(s):  
Cristiano Ialongo ◽  
Antonella Farina ◽  
Raffaella Labriola ◽  
Antonio Angeloni ◽  
Emanuela Anastasi

We read with great interest the paper by Gaudio and colleagues on vitamin D and on the state of patients with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) at the time of admission [...]


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