scholarly journals Concentration and Quantification of SARS-CoV-2 RNA in Wastewater Using Polyethylene Glycol-Based Concentration and qRT-PCR

2021 ◽  
Vol 4 (1) ◽  
pp. 17
Author(s):  
Kata Farkas ◽  
Luke S. Hillary ◽  
Jamie Thorpe ◽  
David I. Walker ◽  
James A. Lowther ◽  
...  
Keyword(s):  
Polyethylene Glycol ◽  
Simple Procedure ◽  
Rna Extraction ◽  
Chemical Properties ◽  
Qrt Pcr ◽  
Reverse Transcription Pcr ◽  
Concentration Method ◽  
Physico Chemical ◽  
One Step ◽  
Wastewater Samples

Wastewater-based epidemiology has become an important tool for the surveillance of SARS-CoV-2 outbreaks. However, the detection of viruses in sewage is challenging and to date there is no standard method available which has been validated for the sensitive detection of SARS-CoV-2. In this paper, we describe a simple concentration method based on polyethylene glycol (PEG) precipitation, followed by RNA extraction and a one-step quantitative reverse transcription PCR (qRT-PCR) for viral detection in wastewater. PEG-based concentration of viruses is a simple procedure which is not limited by the availability of expensive equipment and has reduced risk of disruption to consumable supply chains. The concentration and RNA extraction steps enable 900–1500× concentration of wastewater samples and sufficiently eliminates the majority of organic matter, which could inhibit the subsequent qRT-PCR assay. Due to the high variation in the physico-chemical properties of wastewater samples, we recommend the use of process control viruses to determine the efficiency of each step. This procedure enables the concentration and the extraction the DNA/RNA of different viruses and hence can be used for the surveillance of different viral targets for the comprehensive assessment of viral diseases in a community.

scholarly journals Rapid electrochemical detection of coronavirus SARS-CoV-2

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Thanyarat Chaibun ◽  
Jiratchaya Puenpa ◽  
Tatchanun Ngamdee ◽  
Nimaradee Boonapatcharoen ◽  
Pornpat Athamanolap ◽  
...  
Keyword(s):  
Electrochemical Biosensor ◽  
Rolling Circle Amplification ◽  
Clinical Samples ◽  
Rolling Circle ◽  
Qrt Pcr ◽  
Reverse Transcription Pcr ◽  
Sandwich Hybridization ◽  
Redox Active ◽  
One Step ◽  
The One

AbstractCoronavirus disease 2019 (COVID-19) is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Diagnosis of COVID-19 depends on quantitative reverse transcription PCR (qRT-PCR), which is time-consuming and requires expensive instrumentation. Here, we report an ultrasensitive electrochemical biosensor based on isothermal rolling circle amplification (RCA) for rapid detection of SARS-CoV-2. The assay involves the hybridization of the RCA amplicons with probes that were functionalized with redox active labels that are detectable by an electrochemical biosensor. The one-step sandwich hybridization assay could detect as low as 1 copy/μL of N and S genes, in less than 2 h. Sensor evaluation with 106 clinical samples, including 41 SARS-CoV-2 positive and 9 samples positive for other respiratory viruses, gave a 100% concordance result with qRT-PCR, with complete correlation between the biosensor current signals and quantitation cycle (Cq) values. In summary, this biosensor could be used as an on-site, real-time diagnostic test for COVID-19.

Leveraging Automation toward Development of a High-Throughput Gene Expression Profiling Platform

2020 ◽  
pp. 247255522095659
Author(s):  
Jing Chen ◽  
Alan Futran ◽  
Austin Crithary ◽  
Sha Li ◽  
Alex Wolicki ◽  
...  
Keyword(s):  
Gene Expression ◽  
Drug Discovery ◽  
High Throughput ◽  
Screening Strategy ◽  
Qrt Pcr ◽  
Reverse Transcription Pcr ◽  
Expression Platform ◽  
Program Support ◽  
Drug Discovery Program ◽  
One Step

We previously developed a panel of one-step real-time quantitative reverse transcription PCR (one-step qRT-PCR; hereafter referred to as qRT-PCR) assays to assess compound efficacy. However, these high-cost, conventional qRT-PCR manual assays are not amenable to high-throughput screen (HTS) analysis in a time-sensitive and complex drug discovery process. Here, we report the establishment of an automated gene expression platform using in-house lysis conditions that allows the study of various cell lines, including primary T cells. This process innovation provides the opportunity to perform genotypic profiling in both immunology and oncology therapeutic areas with quantitative studies as part of routine drug discovery program support. This newly instituted platform also enables a panel screening strategy to efficiently connect HTS, lead identification, and lead optimization in parallel.

scholarly journals Industrialisation Scenario at Sreepur of Gazipur, Bangladesh and Physico-chemical Properties of Wastewater Discharged from Industries

2019 ◽  
pp. 1-14 ◽  
Author(s):  
Md. Billal Hossain ◽  
Md. Nurul Islam ◽  
Mohammad Shamsul Alam ◽  
Md. Zakir Hossen
Keyword(s):  
Treatment Plant ◽  
Chemical Properties ◽  
Physicochemical Characteristics ◽  
Mean Value ◽  
Dry Season ◽  
Effluent Treatment ◽  
Industrial Growth ◽  
Physico Chemical ◽  
Textile Dyeing ◽  
Wastewater Samples

This study was conducted to explore the present trend and pattern of industrial growth with spatial distribution of industries and seasonal extent of physicochemical characteristics of wastewater at Sreepur of Gazipur, Bangladesh. The wastewater samples were collected from 5 locations in three seasons viz. pre-monsoon, monsoon and dry season. A total of 120 medium to large industries were surveyed, among those 52 were in red category, 53 were in orange-B category, 13 were in orange-A and only 2 were in green category. In 1995, there were only three industries, which gradually increased to a total of 29 in 2005. Among them, 11 were in red category and 18 were in orange-B category. But from 2006-2010, a total 59 industries were developed and most of them were in red and orange-B categories. Similarly, during the period of 2011 to March 2013, a total 16 industries were developed, among them 8 industries were in red and 3 were in orange-B category. Major types of wastewater discharging industries were textile, dyeing, washing and printing. Among the surveyed industries about 33% didn’t have any effluent treatment plant (ETP). The mean value of pH, EC, DO, BOD, COD and TDS of wastewater were 7.28, 2.64, 1.62 mg L-1, 82.0 mg L-1, 217.31 mg L-1 and 1380 mg L-1, respectively during pre-monsoon; 6.7, 1.15, trace, 8.0 mg L-1, 152.4 mg L-1 and 539.58 mg L-1, respectively during monsoon and 7.7, 1.82, 0.74 mg L-1, 48.8 mg L-1, 204.8 mg L-1 and 993.6 mg L-1, respectively during dry season. Average DO concentrations in all seasons and sites were significantly low, while BOD and COD contents were higher in pre-monsoon and dry seasons than the DoE permissible limit. The study concluded that the area is now a hub of polluting industries which are mostly liable to pollute the surrounding environment.

Designing a Technology of Industrial Sewage Waters Purification from the Spent Polishing Powder Based on Ceric Oxide

2018 ◽  
Vol 284 ◽  
pp. 719-725 ◽  
Author(s):  
T.A. Vasilenko ◽  
A.D. Mishina
Keyword(s):  
Chemical Properties ◽  
Wastewater Purification ◽  
Purification Method ◽  
Ceric Oxide ◽  
Solids Concentration ◽  
Physico Chemical ◽  
Primary Activity ◽  
Wastewater Samples ◽  
Physical And Chemical ◽  
Industrial Sewage

The primary activity of an enterprise, where the wastewater samples have been taken, is applying lacquer coatings to glass and the mirrors production. For washing glass before the application of coatings a suspension of powder based on ceric oxide is prepared, which is used as a polishing solution. The physical and chemical properties of industrial sewage waters, generated after glass polishing, and of the initial and spent polishing powder based on ceric oxide have been researched. The optimal doses of flocculating agents for purifying wastewaters in order to reduce turbidity, suspended solids concentration and dry residue have been determined. After the flocculation the advanced treatment of the water was done, by means of filtering elements with pore size 5 and 1 μm, produced by the Pentek Company. The process scheme of wastewater purification, including the physico-chemical method (flocculation) and mechanical purification method by using bag filters, has been developed.

scholarly journals Rapid Electrochemical Detection of Coronavirus SARS-CoV-2.

2020 ◽  
Author(s):  
Thanyarat Chaibun ◽  
Jiratchaya Puenpa ◽  
Tatchanun Ngamdee ◽  
Nimaradee Boonapatcharoen ◽  
Pornpat Athamanolap ◽  
...  
Keyword(s):  
Electrochemical Biosensor ◽  
Rolling Circle Amplification ◽  
Clinical Samples ◽  
Rolling Circle ◽  
Qrt Pcr ◽  
Reverse Transcription Pcr ◽  
Sandwich Hybridization ◽  
Redox Active ◽  
One Step ◽  
The One

Abstract COVID-19 is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Diagnosis of COVID-19 depends on quantitative reverse transcription PCR (qRT-PCR), which is time-consuming and requires expensive instrumentation. Here, we report an ultrasensitive electrochemical biosensor based on isothermal rolling circle amplification (RCA) for rapid detection of SARS-CoV-2. The assay involves the hybridization of the RCA amplicons with probes that were functionalized with redox active labels that are detectable by an electrochemical biosensor. The one-step sandwich hybridization assay could detect as low as 1 copy/mL of N and S genes, in less than 2 hours. Sensor evaluation with 105 clinical samples, including 40 SARS-CoV-2 positive and 9 samples positive for other respiratory viruses, gave a 100% concordance result with qRT-PCR, with complete correlation between the biosensor current signals and quantitation cycle (Cq) values. In summary, this biosensor could be used as an on-site, real-time diagnostic test for COVID-19.

scholarly journals The physico-chemical properties of 2-methoxyphenoxyacetates of Mn(II), Co(II), Ni(II) and Cu(II)

2018 ◽  
Vol 35 (2) ◽  
pp. 89
Author(s):  
W. Ferenc ◽  
M. Bernat ◽  
J. Sarzyński
Keyword(s):  
Magnetic Moments ◽  
Chemical Properties ◽  
Intermediate Formation ◽  
Bidentate Ligands ◽  
X Ray Diffraction ◽  
Solubility In Water ◽  
Physico Chemical ◽  
One Step ◽  
Thermogravimetric Studies ◽  
Ligand Fields

The complexes of 2-methoxyhenoxyacetates of Mn(II), Co(II), Ni(II) and  Cu(II)with the general formula: M(C9H9O4)3·4H2O, where M(II) = Mn, Co, Ni and Cu have been synthesized and characterized by elemental analysis, IR spectroscopy, magnetic and thermogravimetric studies and also X-ray diffraction measurements. The complexes have colours typical for M(II) ions (Mn(II) - a pale pink, Co(II) - pink, Ni(II) - green, and Cu(II) – blue). The carboxylate group binds as monodentate and bidentate ligands. On heating to 1273K in air thecomplexes decompose in the same way. At first, they dehydrate in one step to anhydrous salts, that next decompose to the oxides of respective metals with the intermediate formation of the oxycarbonates. Their solubility in water at 293K is of the order of 10-5 mol·dm-3. The magnetic moments of analysed complexes were determined in the range of 76-303K. The results reveal them to be high-spin complexes of weak ligand fields.

AN ATTEMPT TO PRODUCE RECOMBINANT HORSE INSULIN

2017 ◽  
Vol 19 (3) ◽  
pp. 3-9
Author(s):  
Batsaikhan D. ◽  
Lkhagvasuren S. ◽  
Munkhtogtokh B. ◽  
Tuvshin B. ◽  
Bayar-Enkh B. ◽  
...  
Keyword(s):  
Recombinant Dna ◽  
Rna Extraction ◽  
Elisa Test ◽  
Recombinant Dna Technology ◽  
Carboxypeptidase B ◽  
Total Rna ◽  
Reverse Transcription Pcr ◽  
Stabilizing Solution ◽  
One Step ◽  
Dna Technology

The present study aimed to produce horse insulin by use of recombinant DNA technology. Because the hormones such as somatotropin, prolactin and leptin, which are practically applicable in veterinary medicine, are also peptides, they can be obtained on the basis of this model technology. To isolate total RNA, horse pancreas samples were collected in RNA stabilizing solution from local abattoirs, sent to the laboratory, and stored the samples in the freezer at -80°C. Molecular biological reagents and kits for total RNA extraction,one step RT (reverse transcription)-PCR reactions, cloning, expression, and purification were used to produce equine proinsulin. Primers were designed from NCBI gene bank data and synthesized in Macrogen Inc, Korea. Obtained equine proinsulin was processed by enzymes trypsin and carboxypeptidase B to form mature insulin. The purity of insulin was assessed by SDS-PAGE. Molecular weight of insulin is approximately 6.0 kDa. The hormone was also measured by ELISA test and it was equal to 55 IU per ml of solution. Finally it can be concluded equine insulin with 55 IU/ml activity can be produced by recombinant DNA technology.

scholarly journals The physico-chemical properties of 4-chlorophenoxyacetates of Mn(II), Co(II), Ni(II) and Cu(II)

2018 ◽  
Vol 33 (1) ◽  
pp. 07
Author(s):  
W. Ferenc ◽  
M. Bernat ◽  
J. Sarzynski ◽  
B. Paszkowska
Keyword(s):  
Magnetic Moments ◽  
Chemical Properties ◽  
Bidentate Ligands ◽  
X Ray Diffraction ◽  
Magnetic Studies ◽  
X Ray ◽  
Physico Chemical ◽  
One Step ◽  
Polycrystalline Solids ◽  
Ligand Fields

The complexes of 4-chlorophenoxyacetates of Mn(II), Co(II), Ni(II) and Cu(II) have been synthesized as polycrystalline solids, and characterized by elemental analysis, spectroscopy, magnetic studies and also by X-ray diffraction and thermogravimetric measurements. The analysed complexes have the following colours: pink for Co(II), green for Ni(II), blue for Cu(II) and a pale pink for Mn(II) compounds. The carboxylate group binds as monodentate and bidentate ligands. On heating to 1173K in air the complexes decompose in several steps. At first, they dehydrate in one step to anhydrous salts, that next decompose to the oxides of respective metals. Their magnetic moments were determined in the range of 76-303K. The results reveal them to be high-spin complexes of weak ligand fields.

scholarly journals Magnetic, spectrochemical and thermal properties of the 2-chloro-5-nitrobenzoates of Co(II), Ni(II) and Cu(II)

2006 ◽  
Vol 71 (8-9) ◽  
pp. 929-937 ◽  
Author(s):  
Wiesława Ferenc ◽  
Agnieszka Walków-Dziewulska ◽  
Beata Cristóvão ◽  
Jan Sarzyñski
Keyword(s):  
Thermal Decomposition ◽  
Thermal Properties ◽  
Metal Ion ◽  
Chemical Properties ◽  
Water Molecules ◽  
Dehydration Process ◽  
Crystallization Water ◽  
Physico Chemical ◽  
One Step ◽  
Hydrated Complexes

Some physico-chemical properties of the 2-chloro-5-nitrobenzoates of Co(II) Ni(II) and Cu(II) were studied. The complexes were obtained as hydrates with a metal ion to ligand ratio of 1:2. All the complexes were polycrystalline compounds. Their colors depended on the nature of the central ion: pink for the Co(II) complex, green for the Ni(II) and blue for the Cu(II) compounds. Their thermal decomposition was studied in the range of 293-523 K because it was found that the 2-chloro-5-nitrobenzoates decomposed explosively on heating in air above 523 K. The hydrated complexes lost crystallization water molecules in one step. From the results, it appeared that no transformation of the nitro group to nitrite occurred during the dehydration process. Their solubilities in water at 293 K were of the order of 10-2 mol dm-3 for the Ni(II) and Co(II) complexes and 10-4 mol dm-3 for that of Cu(II). The magnetic moment values of Co2+, Ni2+ and Cu2+ in the 2-chloro-5-nitrobenzoates, experimentally determined in the range, of 76-303 K ranged from 3.48 to 3.82 ?B for Co(II), from 2.69 to 3.08 ?B for the Ni(II) and from 1.87 to 2.00 ?B for the Cu(II) complex. The 2-chloro-5-nitrobenzoates of Co(II), Ni(II) and Cu(II) followed the Curie-Weiss law.

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