PCR assay Articles
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Boost Assay Performance
Improve SARS-CoV-2 RT-PCR Assays Improve Automated Workflows and Boost Assay Performance Employing a process optimization approach to assay development involves breaking down the assay workflow into smaller modules, such as a mixing step or a reagent addition step, and then measuring and optimizing each module individually using MVS technology. Once the assay is optimized to its fullest extent, ...
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Evaluation of QIAamp DNA mini kit for removing of inhibitors in detection of Cryptosporidium parvum Oocysts in water samples by a nested– PCR assay
In recent years, there has been a dramatic increase in the occurrence of waterborne disease outbreaks caused by the Cryptosporidium parvum, and presence of this protozoan parasite in drinking water is a significant health problem faced by the water industry. A new strategy for detection of Cryptosporidium oocysts in water samples is PCR– based techniques. In this study a nested– PCR assay was ...
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Cyclospora cayetanensis travels in tap water on Italian trainsTap water samples from the toilets of an Italian national railway train were collected over a period of 10 months and tested for the presence of Cyclospora cayetanensis (C. cayetanensis) using EvaGreen® real-time polymerase chain reaction (RT-PCR) assay coupled with high resolution melting (HRM) analysis for protozoan detection and oocyst quantification. C. cayetanensis positive samples ...
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Quantitative characterization of the inhibitory effects of salt, humic acid, and heavy metals on the recovery of waterborne norovirus by electropositive filters
The virus adsorption-elution technique (VIRADEL) using electropositively charged filters is used frequently for recovering enteric viruses from water. The filter-absorbed virus is typically eluted, concentrated, and subsequently detected by culture or molecular methods. Human norovirus (HuNoV), one of the most important waterborne pathogens, cannot be cultivated by conventional culture methods ...
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Multiplex PCR for detection of water-borne bacteria
Microbial water-borne diseases still affect developing countries and are major water quality concerns throughout the world. Routine culture-based methods of identifying bacterial pathogens in water sources are laborious and time-consuming. Recently, the use of molecular techniques such as the polymerase chain reaction (PCR) has provided rapid and highly promising detection methods. In this ...
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Surveillance of
Legionella species in hospital water systems: the significance of detection method for environmental surveillance dataMonitoring of hospital water systems to prevent and control nosocomial legionellosis is important from a public health perspective. This study was conducted to survey the prevalence of Legionella contamination of hospital waters. A total of 44 water samples from the hot-water system of 11 hospitals were tested for Legionella by a culture method and a nested polymerase chain reaction (PCR) assay ...
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Implementation of a process internal control to monitor the performance of real-time PCR assays for the detection of
Vibrio cholerae in waterThe process control utilised in this study was an Escherichia coli (E. coli-GFP) strain carrying a chromosomally integrated copy of the green fluorescent protein (gfp) gene of the jelly fish Aequorea victoria. Application of the process control involved spiking samples with the E. coli-GFP containing cells and detecting gfp using real-time polymerase chain reaction (PCR). The process control ...
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Removal of norovirus from water by coagulation, flocculation and sedimentation processes
In this study, we determined the removal of a prototype human norovirus (Norwalk virus, NV) by bench-scale alum coagulation, flocculation and sedimentation processes using reverse transcriptase polymerase chain reaction (RT-PCR) for norovirus assays. After determining optimum conditions for the coagulation, flocculation and sedimentation processes in terms of turbidity reduction, jar tests were ...
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Rapid and specific detection of
Salmonella in water samples using real-time PCR and High Resolution Melt (HRM) curve analysisA real-time PCR assay combined with a pre-enrichment step for the specific and rapid detection of Salmonella in water samples is described. Following amplification of the invA gene target, High Resolution Melt (HRM) curve analysis was used to discriminate between products formed and to positively identify invA amplification. The real-time PCR assay was evaluated for specificity and sensitivity. ...
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Detection and characterization of human pathogenic viruses circulating in community wastewater using multi target microarrays and polymerase chain reaction
Sewage pollution remains the most significant source of human waterborne pathogens. This study describes the detection and characterization of human enteric viruses in community wastewaters using cell culture coupled with multiple target microarrays (with a total of 780 unique probes targeting 27 different groups of both DNA and RNA viruses) and polymerase chain reaction (PCR) assays. Over a ...
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Ecology of Sulfate-reducing bacteria in an Iron-dominated, mining-impacted freshwater sediment
Received for publication October 31, 2007. A legacy of lead and silver mining in its headwaters left Lake Coeur d'Alene, Idaho with a sediment body that is highly reduced and contains up to 100 g kg–1 iron and a smaller fraction of chemically active sulfide phases. The dynamic character of these sulfides and their importance for the sequestering of contaminating trace elements prompted this study ...
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Detection of
Pseudomonas aeruginosa by PCR in tap-water and bottled mineral water in the Isfahan province of IranThe purpose of this study was to consider the use of a simple polymerase chain reaction (PCR) as an accurate, safe and rapid method to detect Pseudomonas aeruginosa in tap-water and bottled mineral water from Isfahan province, Iran. A total of 224 tap-water and bottled mineral water samples were taken over six months, from July to December 2010. DNA (deoxyribonucleic acid) was extracted from ...
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The detection of
Yersinia enterocolitica in surface water by quantitative PCR amplification of theail andyadA genesYersinia enterocolitica has been detected in surface water, and drinking untreated water is a risk factor for infection. PCR-based methods have been used to detect Y. enterocolitica in various sample types, but quantitative studies have not been conducted in water. In this study, quantitative PCR (qPCR)-based methods targeting the Yersinia virulence genes ail and yadA were used to survey the ...
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SEL, a selective enrichment broth for simultaneous growth of salmonella enterica, escherichia coli o157:h7, and listeria monocytogenes
Multipathogen detection on a single-assay platform not only reduces the cost for testing but also provides data on the presence of pathogens in a single experiment. To achieve this detection, a multipathogen selective enrichment medium is essential to allow the concurrent growth of pathogens. SEL broth was formulated to allow the simultaneous growth of Salmonella enterica, Escherichia coli ...
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Survival of Helicobacter pylori in the wastewater treatment process and the receiving river in Michigan, USA
Contaminated water may play a key role in the transmission of Helicobacter pylori, resulting in gastrointestinal diseases in humans. The wastewater treatment process is an important barrier to control the transmission of H. pylori. However, the presence and viability of H. pylori in the treatment process is not well known. In this paper, the real colony morphology of H. pylori was confirmed ...
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Survey and genetic characterization of wastewater in Tunisia for
Cryptosporidium spp.,Giardia duodenalis, Enterocytozoon bieneusi, Cyclospora cayetanensis andEimeria spp.The microbial diversity of wastewater used for irrigation and fertilization was assessed using specific polymerase chain reaction (PCR) assays to detect and genotype several pathogenic protists including Cryptosporidium spp., Giardia duodenalis, Cyclospora spp., Eimeria spp. and Enterocytozoon bieneusi. A total of 220 wastewater samples (110 raw, 110 treated) and 12 sludge samples were ...
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Spatial distribution of some microbial trophic groups in a plug-flow-type anaerobic bioreactor treating swine manure
Anaerobic digestion of swine manure is carried out by a consortium of microbial species, including volatile fatty acid (VFA) producers, VFA-degraders and methanogens. The distribution of five phylogenetic groups within a plug-flow-type anaerobic bioreactor consisting of eight serially-connected tanks was examined through the sequential digestion of swine manure. Quantification was carried out ...
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Effect of gaseous chlorine dioxide on indoor microbial contaminants
Traditional and modern techniques for bioaerosol enumeration were used to evaluate the relative efficiency of gaseous chlorine dioxide (ClO2) in reducing the indoor microbial contamination under field and laboratory conditions. The field study was performed in a highly microbially contaminated house, which had had an undetected roof leak for an extended period of time and exhibited large areas of ...
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Detection of
Vibrio cholerae andVibrio parahaemolyticus by molecular and culture based methods from source water to household container-stored water at the point-of-use in South African rural communitiesDetection methods for Vibrio cholerae and Vibrio parahaemolyticus which included the culture based approach with polymerase chain reaction (PCR) confirmation, PCR detection without enrichment and PCR with a pre-enrichment were developed and their performance evaluated. PCR assays targeted the SodB (V. cholerae species), Flae (V. parahaemolyticus species), 16S rRNA (Vibrio and Enterobacteriacea ...
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Development and test for long-term stability of a synthetic standard for a quantitative Cryptosporidium parvum LightCycler™ PCR assay
ABSTRACTA recently described quantitative rapid cycle real time PCR (LightCycler™) assay detects Cryptosporidium parvum after in vitro excystation, which is a surrogate marker for the viability of the organisms. In the original assay the quantification standard is a dilution series of C. parvum oocysts with a microscopically determined excystation rate. The need to keep suspensions of viable ...
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