DOI: 10.1016/j.watres.2018.03.023
Scopus记录号: 2-s2.0-85046015496
论文题名: Improved simultaneous quantification of multiple waterborne pathogens and fecal indicator bacteria with the use of a sample process control
作者: Zhang Q. ; Ishii S.
刊名: Water Research
ISSN: 431354
出版年: 2018
卷: 137 起始页码: 193
结束页码: 200
语种: 英语
英文关键词: Fecal indicator bacteria (FIB)
; Microfluidic qPCR
; Pathogens
; Quantitative polymerase chain reaction (qPCR)
; Sample process control (SPC)
; Water quality
Scopus关键词: Chains
; DNA
; Efficiency
; Escherichia coli
; Genes
; Microfluidics
; Molecular biology
; Polymerase chain reaction
; Process control
; Recovery
; Risk assessment
; RNA
; Salmonella
; Water analysis
; Water filtration
; Water quality
; Environmental water samples
; Fecal indicator bacteria
; Gram-negative bacteria
; Listeria monocytogenes
; Quantitative polymerase chain reaction
; Salmonella typhimurium
; Water quality monitoring
; Water-borne pathogens
; Pathogens
; river water
; RNA 23S
; bacterial DNA
; fresh water
; bacterium
; cell
; DNA
; gene
; gene expression
; pathogen
; polymerase chain reaction
; water quality
; waterborne disease
; Article
; bacterial cell
; Campylobacter jejuni
; Campylobacter lari
; Clostridium perfringens
; controlled study
; DNA extraction
; Escherichia coli
; feces microflora
; Gram negative bacterium
; Japan
; Legionella pneumophila
; Listeria monocytogenes
; Mississippi
; nonhuman
; polymerase chain reaction
; priority journal
; process control
; Pseudogulbenkiania
; river
; Salmonella enterica serovar Typhimurium
; Shigella
; transposon
; Vibrio cholerae
; water analysis
; water sampling
; bacterium
; environmental monitoring
; feces
; genetics
; isolation and purification
; microbiology
; Minnesota
; procedures
; water pollutant
; Japan
; Minnesota
; Mississippi River
; United States
; Bacteria (microorganisms)
; Campylobacter jejuni
; Listeria monocytogenes
; Negibacteria
; Salmonella typhimurium
; Bacteria
; DNA, Bacterial
; Environmental Monitoring
; Feces
; Fresh Water
; Japan
; Minnesota
; Polymerase Chain Reaction
; RNA, Ribosomal, 23S
; Water Microbiology
; Water Pollutants
英文摘要: Quantitative polymerase chain reaction (qPCR) is now commonly used to detect fecal indicator bacteria (FIB) as well as pathogens in water samples. However, DNA loss during sample processing can cause underestimation of target genes. In this study, we created a sample process control strain (SPC) by genetically engineering a non-pathogenic, Gram-negative bacterium Pseudogulbenkiania sp. strain NH8B. The SPC strain, named NH8B-1D2, has a kanamycin-resistance gene inserted to one of the 23S rRNA genes. To specifically quantify the SPC strain, a new TaqMan qPCR assay was developed. To obtain the relationship between the DNA recovery efficiencies of various pathogens and those of the SPC strain, known amount of E. coli O157:H7, Salmonella Typhimurium, Campylobacter jejuni, or Listeria monocytogenes cells were co-spiked with the SPC strain to environmental water samples. The DNA recovery efficiencies were calculated by comparing the quantity of bacterial cells inoculated to water samples prior to filtration and DNA extraction, and those measured by qPCR. We then obtained the ratios in the recovery efficiencies between pathogens and SPC strain (RRPATH/SPC). The RRPATH/SPC values obtained using Oono pond water collected in Japan were used as a pathogen-specific constant to estimate the accurate concentrations of pathogens in water samples collected from Mississippi River in Minnesota. Estimated pathogen concentrations were not significantly different from the inoculated pathogen concentration, suggesting our normalization approach is useful to estimate the accurate concentrations of pathogens in environmental water samples. The qPCR assay targeting the SPC strains and FIB were incorporated into the microfluidic qPCR chip format (PBQ chip ver. 2); therefore, we can simultaneously quantify multiple pathogens, FIB, and the SPC strain in high throughput from many water samples. This new tool can be useful for water quality monitoring and risk assessment. © 2018 Elsevier Ltd Citation statistics:
资源类型: 期刊论文
标识符: http://119.78.100.158/handle/2HF3EXSE/112827
Appears in Collections: 气候减缓与适应
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作者单位: BioTechnology Institute, University of Minnesota, St. Paul, MN, United States; Department of Soil, Water, and Climate, University of Minnesota, St. Paul, MN, United States; Division of Environmental Engineering, Hokkaido University, Sapporo, Hokkaido, Japan
Recommended Citation:
Zhang Q.,Ishii S.. Improved simultaneous quantification of multiple waterborne pathogens and fecal indicator bacteria with the use of a sample process control[J]. Water Research,2018-01-01,137