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An efficient method to enhance recovery and detection of SARS-CoV-2 RNA in wastewater

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Journal of environmental Sciences 2023年第8期130卷 139-148页

作者： Teresa Kumblathan Yanming Liu Yuanyuan Qiu Lilly Pang steve e.hrudey X.Chris Le Xing-Fang Li Division of Analytical and environmental Toxicology Department of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonAlbertaT6G 2G3Canada Division of Diagnostic and Applied Microbiology Department of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonT6G 2G3Canada Public Health Laboratory Alberta Precision LaboratoriesEdmontonAlbertaT6G 2G3Canada

Wastewater surveillance(WS)of SARS-CoV-2 currently requires multiple steps and suffers low recoveries and poor ***,we report an improved analytical method with high sensitivity and recovery to quantify SARS-CoV-2 RNA in *** improve the recovery,we concentrated SARS-CoV-2 viral particles and RNA from both the solid and aqueous phases of wastewater using an electronegative membrane(eM).The captured viral particles and RNA on the eM were incubated in our newly developed viral inactivation and RNA preservation(VIP)***,the RNA was concentrated on magnetic beads and inhibitors removed by *** eluting,the RNA on the magnetic beads was directly detected using reverse transcription quantitative polymerase chain reaction(RT-qPCR).Analysis of SARS-CoV-2 pseudovirus(SARS-CoV-2 RNA in a noninfectious viral coat)spiked to wastewater samples showed an improved recovery of 80%.Analysis of 120 wastewater samples collected twice weekly between May 2021 and February 2022 from two wastewater treatment plants showed 100%positive detection,which agreed with the results independently obtained by a provincial public health *** concentrations of SARS-CoV-2 RNA in these wastewater samples ranged from 2.4×10^(2) to 2.9×10^(6) copies per 100 mL of *** method’s capability of detecting trace and diverse concentrations of SARS-CoV-2 in complex wastewater samples is attributed to the enhanced recovery of SARS-CoV-2 RNA and efficient removal of PCR *** improved method for the recovery and detection of viral RNA in wastewater is important for wastewater surveillance,complementing clinical diagnostic tests for public health protection.

关键词： Wastewater Virus Viral RNA environmental monitoring Pathogens Surveillance

Number of COVID-19 cases required in a population to detect SARS-CoV-2 RNA in wastewater in the province of Alberta, Canada:Sensitivity assessment

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Journal of environmental Sciences 2023年第3期125卷 843-850页

作者： Qiaozhi Li Bonita e.Lee Tiejun Gao Yuanyuan Qiu erik ellehoj Jiaao Yu Mathew Diggle Graham Tipples Rasha Maal-Bared Deena Hinshaw Christopher Sikora Nicholas J.Ashbolt James Talbot steve e.hrudey Xiaoli Pang School of Public Health University of AlbertaEdmontonAB T6G 2R3Canada Department of Pediatrics University of AlbertaEdmontonAB T6G 1C9Canada Department of Laboratory Medicine and Pathology University of AlbertaEdmontonAB T6G 2B7Canada University of Alberta Central Receiving EdmontonAlbertaT6G 2R3Canada Provincial Laboratory for Public Health EdmontonAlbertaCanada ePCOR EdmontonAB T5H 0E8Canada Department of Medicine University of AlbertaEdmontonAB T6G 2G3Canada Faculty of Science and engineering Southern Cross UniversityEast Lismore NSW 2480Australia

With a unique and large size of testing results of 1,842 samples collected from 12 wastewater treatment plants (WWTP) for 14 months through from low to high prevalence of COVID-19,the sensitivity of RT-qPCR detection of SARS-CoV-2 RNA in wastewater that correspond to the communities was computed by using Probit *** study determined the number of new COVID-19 cases per 100,000 population required to detect SARS-CoV-2RNA in wastewater at defined probabilities and provided an evidence-based framework of wastewater-based epidemiology surveillance (WBe).Input data were positive and negative test results of SARS-CoV-2 RNA in wastewater samples and the corresponding new COVID-19 case rates per 100,000 population served by each *** analyses determined that RT-qPCR-based SARS-CoV-2 RNA detection threshold at 50%,80%and 99%probability required a median of 8 (range:4-19),18 (9-43),and 38 (17-97) of new COVID-19 cases/100,000,***,the positive detection rate at 50%,80%and 99%probability were 0.01%,0.02%,and 0.04%averagely for new cases in the *** study improves understanding of the performance of WBe SARS-CoV-2 RNA detection using the large datasets and prolonged study *** COVID-19 burden at a community level that would result in a positive detection of SARS-CoV-2 in wastewater is critical to support WBe application as a supplementary warning/monitoring system for COVID-19 prevention and control.

关键词： SARS-CoV-2 COVID-19 cases Wastewater Probit analysis Probability Detection sensitivity

Nontargeted identification of peptides and disinfection byproducts in water

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Journal of environmental Sciences 2016年第4期28卷 259-266页

作者： Yanan Tang Ying Xu Feng Li Lindsay Jmaiff steve e.hrudey Xing-Fang Li Division of Analytical and environmental Toxicology Department of Laboratory Medicine and PathologyUniversity of Alberta Department of Computer Science University of Alberta

A broad range of organic compounds are known to exist in drinking water sources and serve as precursors of disinfection byproducts（DBPs）.epidemiological findings of an association of increased risk of bladder cancer with the consumption of chlorinated water has resulted in health concerns about *** are thought to be an important category of DBP precursors in ***,little is known about the actual presence of peptides and their DBPs in drinking water because of their high sample complexity and low *** address this challenge and identify peptides and non-chlorinated/chlorinated peptide DBPs from large sets of organic compounds in water,we developed a novel high throughput analysis strategy,which integrated multiple solid phase extraction（SPe）,high performance liquid chromatography（HPLC）separation,and non-target identification using precursor ion exclusion（PIe）high resolution mass spectrometry（MS）.After MS analysis,structures of candidate compounds,particularly peptides,were obtained by searching against the Human Metabolome Database（HMDB）.Using this strategy,we successfully detected 625 peptides（out of 17,205 putative compounds）and 617 peptides（out of 13,297）respectively in source and finished water *** source and finished water samples had 501 peptides and amino acids in *** remaining 116 peptides and amino acids were unique to the finished *** a subset of 30 putative compounds for which standards were available,25 were confirmed using HPLC-MS *** analyzing the peptides identified in source and finished water,we successfully confirmed three disinfection reaction pathways that convert peptides into toxic DBPs.

关键词： Nontargeted detection Water analysis Mass spectrometry Peptide Disinfection byproducts

N-Nitrosamine formation from chloramination of two common ionic liquids

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Journal of environmental Sciences 2020年第1期32卷 341-348页

作者： Ian J.Vander Meulen Ping Jiang Di Wu steve e.hrudey Xing-Fang Li Division of Analytical and environmental Toxicology Department of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of Alberta

Ionic liquids(ILs)are a class of solvents increasingly used as"green chemicals."Widespread applications of ILs have led to concerns about their accidental entry to the *** have been assessed for some environmental impacts;however,little has been done to characterize their potential impacts on drinking water if ILs accidentally enter surface *** cations are often aromatic or alkyl quaternary amines that resemble structures of previously confirmed N-nitrosamine(NA)***,this study has evaluated two common ILs,1-ethyl-3-methylimidazolium bromide(eMIm Br)and 1-ethyl-1-methylpyrrolidinium bromide(eMPyr Br),for their NA formation *** IL species was reacted with pre-formed monochloramine under various laboratory *** reaction mixtures were extracted using liquid–liquid extraction and analyzed for NAs using high performance liquid chromatography tandem mass *** low concentration of IL(250μmol/L),the yields of NAs(NMeA or NPyr)increased with increasing doses of monochloramine from both IL *** total NA yield was as high as 2.5±0.3 ng/mg from eMIm Br,and as high as 8.6±0.8 ng/mg from eMPyr *** high concentration of IL(5 mmol/L),the NA yield reached a maximum at 2.5 mmol/L NH2Cl,and then decreased with subsequent increases in the reactant concentrations,demonstrating ILs’solvent *** study re-emphasizes the importance of preventing discharge of ILs to water bodies to prevent secondary impacts on drinking water.

关键词： Ionic liquids N-nitrosamine Liquid–liquid extraction HPLC–MS/MS DBPs

Quantification and Differentiation of SARS-CoV‑2 Variants in Wastewater for Surveillance

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environment & Health 2023年第3期1卷 203-213页

作者： Teresa Kumblathan Yanming Liu Xiaoli Pang steve e.hrudey XChris Le Xing-Fang Li Division of Analytical and environmental Toxicology Department of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonAlbertaCanada T6G 2G3 Division of Diagnostic and Applied Microbiology Department of Laboratory Medicine and PathologyFaculty of Medicine and DentistryUniversity of AlbertaEdmontonAlbertaCanada T6G 2B7 Public Health Laboratory Alberta Precision LaboratoriesEdmontonAlbertaCanada T6G 2J2

Wastewater surveillance plays an important role in the monitoring of infections of SARS-CoV-2 at the community *** report here the determination of SARS-CoV-2 and differentiation of its variants of concern in 294 wastewater samples collected from two major Canadian cities from May 2021 to March *** overall method of analysis involved extraction of the virus and viral components using electronegative membranes,in situ stabilization and concentration of the viral RNA onto magnetic beads,and direct analysis of the viral RNA on the magnetic *** reverse transcription quantitative polymerase chain reaction(RT-qPCR)assays,targeting specific and naturally selected mutations in SARS-CoV-2,enabled detection and differentiation of the Alpha,Beta,Gamma,Delta,and Omicron *** Omicron triplex RT-qPCR assay targeting three mutations,HV 69−70 deletion,K417N,and L452R,was able to detect and differentiate the Omicron BA.1/BA.3,BA.2/XBB,and BA.4/*** assay had efficiencies of 90−104%for all three mutation targets and a limit of detection of 28 RNA copies per *** of 294 wastewater samples collected over a two-year span showed the concentrations and trends of Alpha,Beta,Gamma,Delta,and Omicron variants as they emerge in two major Canadian cities participating in the wastewater surveillance *** trends of specific variants were consistent with clinical reports for the same *** the beginning of each wave,the corresponding variants were detectable in *** example,RNA concentrations of the BA.2 variant were as high as 10^(4)copies per 100 mL of wastewater collected in January 2022,when approximately only 50−60 clinical cases of BA.2 infection were reported in *** results show that the strategy and highly sensitive assays for the variants of concern in wastewater are potentially useful for the detection of newly emerging SARS-CoV-2 variants and other viruses for future community biomonitoring.

关键词： COVID-19 SARS-CoV-2 variants of concern Omicron subvariants multiplex RT-qPCR wastewater surveillance

Comparison of approaches to quantify SARS-CoV-2 in wastewater using RT-qPCR: Results and implications from a collaborative inter-laboratory study in Canada

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Journal of environmental Sciences 2021年第9期33卷 218-229页

作者： Alex H.S.Chik Melissa B.Glier Mark Servos Chand S.Mangat Xiao-Li Pang Yuanyuan Qiu Patrick M.D’Aoust Jean-Baptiste Burnet Robert Delatolla Sarah Dorner Qiudi Geng John P.Giesy Jr Robert Mike McKay Michael R.Mulvey Natalie Prystajecky Nivetha Srikanthan Yuwei Xie Bernadette Conant steve e.hrudey Canadian SARS-CoV-Inter-Laboratory Consortium Consultant to Canadian Water Network Inc. KitchenerCanada Presently at Ontario Clean Water Agency MississaugaCanada environmental Microbiology BC Centre for Disease ControlVancouverCanada Department of Biology University of WaterlooWaterlooCanada National Microbiology Laboratory Public Health Agency of CanadaWinnipegCanada Public Health Laboratory Alberta Precision LaboratoryEdmontonCanada Department of Laboratory Medicine&Pathology Faculty of Medicine and DentistryUniversity of AlbertaEdmontonAB T6G 2G3Canada Faculty of engineering University of OttawaOttawaCanada Département des génies civil géologique et des minesPolytechnique MontréalMontréalCanada Veterinary Biomedical Sciences University of SaskatchewanSaskatoonCanada Toxicology Centre University of SaskatchewanSaskatoonCanada Great Lakes Institute for environmental Research University of WindsorWindsorCanada Department of Medical Microbiology University of ManitobaWinnipegCanada Department of Pathology and Laboratory Medicine University of British ColumbiaVancouverCanada Canadian Water Network Inc. WaterlooCanada Group author details below

Detection of SARS-CoV-2 RNA in wastewater is a promising tool for informing public health decisions during the COVID-19 ***,approaches for its analysis by use of reverse transcription quantitative polymerase chain reaction(RT-q PCR)are still far from standardized *** characterize inter-and intra-laboratory variability among results when using various methods deployed across Canada,aliquots from a real wastewater sample were spiked with surrogates of SARS-CoV-2(gamma-radiation inactivated SARS-CoV-2 and human coronavirus strain 229e[HCoV-229e])at low and high levels then provided"blind"to eight *** estimates reported by individual laboratories were consistently within a 1.0-log_(10) range for aliquots of the same spiked *** laboratories distinguished between low-and high-spikes for both *** expected,greater variability was observed in the results amongst laboratories than within individual laboratories,but SARS-CoV-2 RNA concentration estimates for each spiked condition remained mostly within 1.0-log_(10) *** no-spike wastewater aliquots provided yielded non-detects or trace levels(ene copies/mL)of SARS-CoV-2 *** appear linked to methods that included or focused on the solids fraction of the wastewater matrix and might represent in-situ SARS-CoV-2 to the wastewater ***-229e RNA was not detected in the no-spike ***,all methods yielded comparable results at the conditions *** behavior of SARS-CoV-2 and spiked surrogates in wastewater should be considered to evaluate method effectiveness.A consistent method and laboratory to explore wastewater SARS-CoV-2 temporal trends for a given system,with appropriate quality control protocols and documented in adequate detail should succeed.

关键词： COVID-19 Wastewater surveillance Public health Quality assurance Quality control

Confirmation of a Sub-Saturn-size Transiting exoplanet Orbiting a G Dwarf:TOI-1194 b and a Very Low Mass Companion Star: TOI-1251 B from TeSS

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Research in Astronomy and Astrophysics 2024年第3期24卷 129-143页

作者： Jia-Qi Wang Xiao-Jun Jiang Jie Zheng Hanna Kellermann Arno Riffeser Liang Wang Karen A.Collins Allyson Bieryla Lars A.Buchhave steve B.Howell elise Furlan eric Girardin Joao Gregorio eric Jensen Felipe Murgas Mesut Yilmaz Sam Quinn Xing Gao Ruo-Yu Zhou Frank Grupp Hui-Juan Wang CAS Key Laboratory of Optical Astronomy National Astronomical ObservatoriesChinese Academy Of SciencesBeijing 10010China University of Chinese Academy of Sciences Beijing 100049China University Observatory Munich Scheinertrsse 1.D-81679 MunichGemany Max Planck Institute for extraterrestrial Physics Giessenbachstr.1D-85748 GarchingGermany National Astronomical Observatories/Nanjing Institute of Astronomical Optics&Technology Chinese Academy of SciencesNanjing210042China CAS Key Laboratory of Astronomical Optics&Technology Nanjing Institute of Astronomical Optics&TechnologyNanjing 210042China Center for Astrophysics Harvard&Smithsonian60 Garden Street.CambridgeMA 02138USA National Space Institute Technical University of DenmarkElektrovejDK-2800 KgsLyngbyDenmark NASA Ames Research Center Moffett FieldCA 94035USA NASA exoplanet Science Institute Caltech/IPACMail Code 100-221200 E.California Blvd.PasadenaCA91125USA Grand Pra Observatory Switzerland Department of Physics Washington UniversitySt.LouisMO 63130.USA Department of Physics and Astronomy Swarthmore CollegeSwarthmorePA 19081USA Instituto de Astrofísica de Canarias(IAC) E-38205 La Laguna.TenerifeSpain Departamento de Astrofísica Universidad de La LagunaSpain Dept.of Astronomy and Space Sciences Ankara UniversityTandogan 06100AnkaraTurkey Xinjiang Astronomical Observatory Chinese Academy of SciencesUrumqi 830011China Urumqi No.1 Senior High School Urumqi 830002China Lijiang Gemini Observatory Lijiang 674199China

We report the confirmation of a sub-Saturn-size exoplanet,TOI-1194 b,with a mass of about 0.456+0.055-0.051M_(J),and a very low mass companion star with a mass of about 96.5±1.5 MJ,TOI-1251 *** candidates provided by the Transiting exoplanet Survey Satellite(TeSS)are suitable for further follow-up observations by ground-based telescopes with small and medium *** analysis is performed based on data from several telescopes worldwide,including telescopes in the Sino-German multiband photometric campaign,which aimed at confirming TeSS Objects of Interest(TOIs)using ground-based small-aperture and medium-aperture telescopes,especially for long-period ***-1194 b is confirmed based on the consistent periodic transit depths from the multiband photometric *** measure an orbital period of 2.310644±0.000001 days,the radius is 0.767+0.045-0.041RJ and the amplitude of the RV curve is 69.4_(-7.3)^(+7.9)m s^(-1).TOI-1251 B is confirmed based on the multiband photometric and high-resolution spectroscopic data,whose orbital period is 5.963054+0.000002-0.000001days,radius is 0.947+0.035-0.033 R_(J) and amplitude of the RV curve is 9849_(-40)^(+42)ms^(-1).

关键词： planets and satellites fundamental parameters-planets and satellites gaseous planets-stars fundamental parameters-stars low-mass-methods data analysis-techniques photometric-techniques spectroscopic

exome sequencing identifies a likely causative variant in 53%of families with ciliopathy-related features on renal ultrasound after excluding NPHP1 deletions

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Genes & Diseases 2024年第5期11卷 40-43页

作者： Konstantin Deutsch Verena Klambt Thomas M.Kitzler Tilman Jobst-Schwan Ronen Schneider Florian Buerger steve Seltzsam Sherif el Desoky Jameela A.Kari Farkhanda Hafeez Maria Szczepanska Loai A.eid Hazem S.Awad Muna Al-Saffar Neveen A.Soliman Velibor Tasic Camille Nicolas-Frank Kirollos Yousef Luca M.Schierbaum Sophia Schneider Abdul Halawi Izzeldin elmubarak Katharina Lemberg Shirlee Shril Shrikant M.Mane Nancy Rodig Friedhelm Hildebrandt Division of Nephrology Department of PediatricsBoston Children’s HospitalHarvard Medical SchoolBostonMA 02215USA Department of Pediatric Gastroenterology Nephrology and Metabolic DiseasesCharite Universitatsmedizin BerlinBerlin 13353Germany Berlin Institute of Health at Charitee Universitatsmedizin Berlin BIH Biomedical Innovation AcademyBIH Charite Clinician Scientist ProgramBerlin 10178Germany Department of Nephrology and Hypertension FriedrichAlexander-Universitat Erlangen-NurnbergErlangen 91054Germany Department of Pediatrics Faculty of Medicine King Abdulaziz UniversityPediatric Nephrology Center of ExcellenceKing Abdulaziz University HospitalJeddah 21589Saudi Arabia Department of Pediatric Nephrology The Children’s Hospital and Institute of Child HealthLahore 54000Pakistan Department of Pediatrics Faculty of Medical Sciences in ZabrzeMedical University of Silesia in KatowiceKatowice 40-752Poland Dubai Hospital and Al-Jalila Children’s Specialty Hospital Kidney Center of ExcellenceDubai 4545United Arab Emirates Department of Genetics and Genomics UAE UniversityAbu Dhabi 15551United Arab Emirates Department of Pediatrics Division of Genetics and GenomicsBoston Children’s HospitalHarvard Medical SchoolBostonMA 02215USA Department of Pediatrics Center of Pediatric Nephrology and TransplantationKasr Al Ainy School of MedicineCairo UniversityCairo 11562Egypt egypt Center for Research and Regenerative Medicine(eCRRM) Cairo 11511Egypt Medical Faculty Skopje University Children’s HospitalSkopje 1000North Macedonia Department of Genetics Yale University School of MedicineNew HavenCT 06520USA

Nephronophthisis-related ciliopathies(NPHP-RC)represent one of the most common causes of chronic kidney disease in the first three decades of life and are characterized by a broad genetic and clinical heterogeneity.1 To date,more than 90 genes have been identified that cause autosomalrecessive NPHP-RC if mutated,accounting for up to 60%of cases.1 Among these,homozygous deletions of NPHP1 are the most common *** genes localize to primary cilia,basal bodies,or the centrosome and lead to a primary ciliary disruption if mutated,thereby causing a broad phenotypical spectrum.1 Patients that suffer from NPHP-RC can either have an isolated renal phenotype,such as polyuria,polydipsia,decreased urinary concentration ability,and secondary enuresis due to loss of tubular function,or present with extrarenal symptoms including retinal degeneration,cerebellar vermis hypoplasia,or hepatic *** are often diagnosed in early adolescence,reaching end-stage renal disease before 25 years of age,but early onset and rapidly progressive forms of NPHP-RC also exist.1 Renal ultrasound indicates kidneys of normal or reduced renal length with increased echogenicity and corticomedullary cysts.

关键词： degeneration urinary likely

Fourier coefficients of restrictions of eigenfunctions

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Science China Mathematics 2023年第8期66卷 1849-1878页

作者： emmett L.Wyman Yakun Xi steve Zelditch Department of Mathematics University of RochesterRochesterNY 14627USA School of Mathematical Sciences Zhejiang UniversityHangzhou 310027China Department of Mathematics Northwestern UniversityEvanstonIL 60208USA

Let{e_(j)}be an orthonormal basis of Laplace eigenfunctions of a compact Riemannian manifold(M,g).Let H■M be a submanifold and{ψ_(k)}be an orthonormal basis of Laplace eigenfunctions of H with the induced *** obtain joint asymptotics for the Fourier coefficientse_(j),ψ_(k)>L^(2)(H)=∫He_(j),ψ_(k)dV_(H)of restrictionsγHe_(j)of e_(j)to *** particular,we obtain asymptotics for the sums of the norm-squares of the Fourier coefficients over the joint spectrum{(μ_(k),λ_(j))}^(∞)_(j,k-0)of the(square roots of the)Laplacian△_(M)on M and the Laplacian△_(H)on H in a family of suitably‘thick'regions in R^(2).Thick regions include(1)the truncated coneμ_(k)/λ_(j)∈[a,b]■(0,1)andλ_(j)≤λ,and(2)the slowly thickening strip|μ_(k)-cλ_(j)|≤w(λ)andλ_(j)≤λ,where w(λ)is monotonic and 1■w(λ)≤λ^(1/2).Key tools for obtaining the asymptotics include the composition calculus of Fourier integral operators and a new multidimensional Tauberian theorem.

关键词： eigenfunctions period integrals Kuznecov formula