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At least 721 records · Page 40Linked to original sources

PubMed1991

An antigen-capture ELISA detects pestivirus antigens in blood and tissues of immunotolerant carrier cattle.

An antigen-capture ELISA was developed for the detection of pestivirus-specific antigens in peripheral blood leucocytes (PBLs), blood clots and tissue samples of immunotolerant cattle persistently infected with virus. The ELISA demonstrated complete agreement with conventional virus isolation procedures undertaken on specimens from a total of 58 carrier animals and 360 uninfected animals. The technique is based on capturing antigen with a high-titred goat polyclonal antiserum and detecting the bound antigen with a combination of 3 broadly-reactive monoclonal antibodies. Increased sensitivity was obtained with the use of an avidin-biotin complex (ABC) amplification method. On average, ELISA optical densities (ODs) for PBL and blood clot samples derived from carrier animals were 1.53 and 0.95, respectively, while uninfected animals had corresponding values of less than 0.15 for all blood samples. Tissue samples from carrier cattle had OD values ranging from an average of 0.95 for liver to 1.77 for spleen, with negative values for all tissues again averaging less than 0.20. Signal-to-noise (S/N) ratios calculated from the ELISA OD readings for carrier cattle showed an average of 15.6 for blood samples and 16.4 for tissues. In contrast, all samples from negative cattle had S/N ratios less than 2.0. The antigen-capture ELISA has been validated on field samples and is suitable for routine diagnostic and certification testing.

Animals↗

PubMed1992

Rapid detection of equine herpesvirus type-1 antigens in nasal swab specimens using an antigen capture enzyme-linked immunosorbent assay.

An antigen capture enzyme-linked immunosorbent assay (ELISA) was developed for the detection of equine herpesvirus type-1 (EHV-1) antigens in nasal swab specimens. The test was designed as a solid phase, amplified sandwich assay in which an EHV-1 specific monoclonal antibody was used to capture virus antigen and polyclonal antisera used to detect antigen bound to the test plates. Eight monoclonal antibodies were tested for their ability to capture virus antigen and one was selected for routine use. The sensitivity and specificity of the ELISA was compared with that of virus isolation using swabs from ponies which were experimentally infected with EHV-1. Of 72 nasal swabs collected, 32 were found to be positive by both virus isolation (VI) and ELISA, a further 15 samples were positive by VI alone, but none of the samples were positive by ELISA and negative by VI. This yielded an overall assay sensitivity of 68% and specificity of 100%. The assay proved useful for diagnosis since virus antigen was detected during the first four days post-infection which corresponded to the acute phase of disease when some clinical symptoms were apparent. In addition, the assay could be completed within one day when antibody coated plates were available.

Animals↗

PubMed1995

A synthetic parvovirus B19 capsid protein can replace viral antigen in antibody-capture enzyme immunoassays.

To establish a renewable source of parvovirus B19 antigens for diagnostic tests, gene sequences for the viral capsid proteins, VP1 and VP2, were cloned into baculovirus expression vectors and the recombinant viruses used to infect Sf9 insect cells. Cell lysates examined by immunoblotting demonstrated reactive proteins corresponding to the expected sizes of native VP1 (83 kDa) and VP2 (58 kDa). The VP2 protein was produced efficiently in quantity and self-assembled into empty capsids as shown by density equilibration in a CsCl step gradient. The VP2 protein was purified and used as an antigen in antibody-capture enzyme immunoassays for the detection of B19 IgG and IgM antibodies. Compared to a standard antibody-capture EIA based on whole viral antigen, the VP2-EIA gave a sensitivity of 100% and specificity of 97% in detection of B19 IgM in 138 patients suspected of B19 infection. No IgM-positive specimens were missed. IgG detection yielded a sensitivity of 100% and specificity of 96% in the same population. Recombinant VP2 capsid proteins expressed in baculovirus-infected insect cells can substitute for serum-derived B19 virus in standard antibody-capture EIA for the detection of B19 IgG and IgM with comparable results.

Adult↗

PubMed1996

Development and application of HHV-6 antigen capture assay for the detection of HHV-6 infections.

An HHV-6 antigen capture assay measuring gp116/64/54 antigen was developed. This ELISA is specific for HHV-6 Variants A and B, does not cross react with other human herpesviruses, is sensitive, stable, quantitative, and can detect antigen in body fluids and cell cultures. Relative to virus isolation or techniques for measuring HHV-6 nucleic acids, the assay is much simpler and less expensive to perform. Plasmas/sera (413) obtained from healthy donors, children with Exanthem subitum, febrile illnesses, patients with Chronic Fatigue Syndrome, and AIDS patients tested by antigen capture assay demonstrated that the assay is useful in clinical laboratory settings. The capture assay can also be used to monitor cell cultures for virus isolation, production, quantitation, and antiviral agent screening.

AIDS-Related Opportunistic Infections↗

PubMed1985

Changes in blood metabolites following stress from capture and handling of the marine teleost Girella tricuspidata.

Whole blood nucleoside triphosphate (NTP) and lactate in the parore (Girella tricuspidata, Fam: Kyphosidae) were monitored over a period of 12 hr following capture by gill net. An increase in NTP during the post-capture recovery period was mainly attributable to a significant rise (P less than 0.05) in the NTP component guanosine triphosphate (GTP). The rise in GTP levels correlated with the decline in blood lactate (r = -0.72) accumulated during the period of capture stress. It is suggested that metabolism of lactate via the Krebs cycle may be responsible for the rise in GTP.

Adenosine Triphosphate↗

PubMed1996

Perception of detail and greyscale range in X-ray fluoroscopy images captured with a personal computer and frame-grabber.

OBJECTIVE: To assess the dynamic range of radiological images captured with a personal computer frame-grabbing system coupled to an X-ray fluoroscopy machine. METHODS: A 386DX-40 MHz, IBM compatible, computer with an SVGA monochrome graphics subsystem and a 387 co-processor, installed with a Screen Machine frame-grabber and controlled by a program specially written was used. Various systems were examined and the observer's perceptions of the results assessed. RESULTS: The dynamic range available to an ordinary X-ray fluoroscopy system was found to be restricted to about 750 mV. Similar measurements showed that the dynamic range was always restricted to 3/4-1/2 of the full available signal because of a high value of the dark voltage of the TV camera's target on all seven systems measured. The dynamic range of the computer-frame grabber system was found to be significantly wider than the Image Intensifier-TV camera chain but, surprisingly, it was affected by the type of file format used for image storing on disk. Clinical images from a barium meal examination as well as CT images captured after optimisation of the frame-grabber were found to contain large quantities of noise in the first two least significant bit planes making them redundant and limiting the grey levels needed for image display to less than 64. This number was also less than the 80 grey levels that could be discriminated by the human eye on the computer monitor. CONCLUSIONS: It was concluded that 6 bit digitisation would have been sufficient for image capture. The advantages of the wider dynamic range of the frame-grabber and the processing capabilities of the computer were tested for the possibility of improving the perception of detail. However, the results were negative. The limiting spatial resolution measured with a variable density bar pattern at all magnifications was about 0.4 lp/mm lower from that measured directly on the fluoroscopic screen. A detail perception test had the same result. The perception success was significantly lower with the digital images at all but the highest of the exposure rates and despite the use of image processing filters.

Artifacts↗

PubMed1990

Multiplex PCR amplification and immobilized capture probes for detection of bacterial pathogens and indicators in water.

Detection of pathogens (Legionella species) and indicator bacteria (coliform bacteria) was achieved by multiplex (simultaneous) PCR amplification of diagnostic gene sequences and by hybridization to immobilized poly-dT-tailed capture probes using a dot- or slot-blot approach. Complex manipulations of primer concentrations and staggered additions of primers were required in order to achieve equal amplification of multiple genes. Multiplex PCR amplification of two different Legionella genes, one specific for L. pneumophila (mip) and the other for the genus Legionella (5S rRNA), was achieved by staggered amplification. Multiplex PCR amplification using differing amounts of primers specific for lacZ and lamB genes permitted the detection of coliform bacteria and those associated with human faecal contamination, including the indicator bacterial species E. coli and enteric pathogens Salmonella and Shigella. Hybridization of biotin-labelled amplified DNA, in which the biotin was incorporated during PCR amplification from biotinylated-dUTP, to immobilized 400-dT-tailed capture probes permitted specific and sensitive detection of target gene sequences. The sensitivity of colorimetric detection achieved by PCR amplification of target DNA was at a level equivalent to 1-2 bacterial cells, which is the same level of sensitivity obtained with radioactive detection. The simultaneous amplification of several genes and hybridization to immobilized capture probes with colorimetric detection is an effective, efficient and rapid detection method for various human bacterial pathogens.

Bacteriological Techniques↗

PubMed1988

An application of capture-recapture methods to the estimation of completeness of cancer registration.

Completeness of cancer registration has not been consistently ascertained across different registries. This report describes how capture-recapture methods have been used to estimate completeness at the Ontario Cancer Registry. The method was applied in two fashions; first, using three data sources in a modeling approach: and second, using two data sources and standard, simple capture-recapture methods. The modeling approach is more flexible, since several variables that influence cancer registration can be considered and can be used to identify reporting patterns of different data sources. In the present analysis, estimates of completeness of the registry as a whole were remarkably similar using either two or three data sources, and site-specific comparisons differed by at most 7%. Because of the advantages of capture-recapture methods-estimation of level of completeness, possible comparability of estimates across different registries, and versatility to consider other determinants of cancer registration-a plea for greater use of these methods in cancer registration is made.

Data Collection↗

PubMed1994

A shortened dengue IgM capture ELISA using simultaneous incubation of antigen and peroxidase-labeled monoclonal antibody.

A shortened IgM capture ELISA for the detection of dengue IgM antibodies using simultaneous incubation of antigen and peroxidase-labeled monoclonal antibody was described. The shortened two-step assay was compared with the four-step IgM capture ELISA on sera from dengue patients confirmed by the hemagglutination inhibition (HI) test. When paired acute and convalescent sera were tested, the shortened ELISA showed 100% agreement with HI results. It detected dengue IgM antibodies in the acute sera of 66% of patients with a primary dengue infection, 60% of patients with a secondary infection, and 98% of patients with a presumptive secondary infection. When the results of 151 dengue patients were combined, 75% of the acute sera were positive by the shortened IgM capture ELISA.

Journal Article↗

PubMed1994

Rapid isolation of cosmid insert DNA by triple-helix-mediated affinity capture.

A simple and rapid method for the isolation of cosmid insert DNA was developed based on triple-helix-mediated affinity capture (TAC). A modified cosmid was constructed from the SuperCos 1 cosmid vector by flanking the cloning site with two homopurine-homopyrimidine triple-helix-forming sequences. The cosmid DNA is digested with NotI restriction enzyme to release the insert DNA. The NotI-digested cosmid DNA is then combined with a biotinylated homopyrimidine oligonucleotide in an acidic buffer solution to form a triple-helix complex. The triple-helix complex is captured with streptavidin-coated magnetic beads. Insert DNA is eluted by adding a pH 9 buffered solution to the captured complex. The purified insert DNA is recovered with a yield of up to 95% and a purity of at least 95%. The isolated insert DNA was directly digested with CviJI restriction endonuclease to generate random fragments for shotgun sequencing.

Base Sequence↗

PubMed2005

Estimating crashes involving heavy vehicles in Western Australia, 1999-2000: a capture-recapture method.

A two-sample exploratory study of police and hospital records was undertaken to estimate the number of fatalities and serious injuries for heavy vehicle drivers involved in a crash in Western Australia. The capture-recapture method was used to assess differences and similarities in characteristics of heavy vehicle drivers from both sources. Each heavy vehicle driver involved in a crash from the police report was matched against the heavy vehicle driver's hospitalisation record from the Hospital Morbidity Data System, with surname, initials, date of birth, gender, date of crash, road user type and vehicle type as matching fields. The estimated number of fatalities and serious injuries to heavy vehicle drivers from 1st July 1999 to 31st December 2000 was 5 and 59, respectively, which was 25 and 31% higher based on the capture-recapture methodology than the aggregated (non-overlapping) total officially reported to the police and hospitals. No significant age difference (p>0.05) was found for drivers involved in a heavy vehicle crash between the two sources (37 years versus 40 year of age). However, female heavy vehicle drivers were over-represented in the hospital records (11%) compared to the police records (1%). The capture-recapture approach is useful for evaluating the completeness of data sources and identifying biases within datasets. The underestimation of heavy vehicle drivers seriously injured and killed has important implications for heavy vehicle safety management and resource allocation in Western Australia.

Accidents, Traffic↗

PubMed2006

Estimating non-fatal road casualties in a large French county, using the capture-recapture method.

INTRODUCTION: Non-fatal road casualties are under-reported, with official figures coming from the police. In the French Rhône county, a road trauma registry constitutes a second source of reporting but its completeness needs to be assessed. We also wish to estimate an incidence rate of non-fatal road casualties that is corrected for under-count. METHODS: Having two sources of reporting available, we can apply the capture-recapture method under certain conditions. To take into account different reporting probabilities among casualties, the capture-recapture analysis is stratified according to injury severity (New Injury Severity Score=NISS), road user type and human third party. To evaluate the sensitivity of the capture-recapture estimate on the number of matched casualties between the police file and the registry, three scenarios of record-linkage are considered. RESULTS: For serious (NISS 9+) non-fatal road casualties, the police ascertainment rate is at most 57%, the registry ascertainment rate is at most 87%, and the aggregate ascertainment rate is at most 95%. The ascertainment-corrected incidence rate for serious (NISS 9+) non-fatal road casualties is at least 65 per 100,000. CONCLUSION: The degree of completeness of the registry for serious casualties is rather high, though not satisfactory.

Accidents, Traffic↗

PubMed2005

Magnetic colloids for the generic capture of viruses.

In molecular biology or in the diagnostic field, a strong interest has been devoted to the use of magnetic nanoparticles as an efficient tool for a fast and easy biomolecule extraction. Various methods allow the synthesis of numerous types of magnetic particles, but very few of them can be adapted to biological protocols. For biological tests based on the final detection of nucleic acids, the major difficulties in the choice of these magnetic beads are their fast magnetic separation, requiring a high magnetic oxide content and overall a good compatibility with enzymes used for nucleic acid amplification. In this study, magnetic colloids were obtained from oil in water (o/w) magnetic emulsion by a two-step polymer immobilization procedure. First, a polycation, poly(ethyleneimine), was adsorbed to ensure charge inversion of the emulsion droplets. Then poly(maleic anhydride-co-methyl vinyl ether) was either chemically grafted or adsorbed if under its hydrolyzed form. The obtained colloids were characterized in terms of colloidal stability, surface charge density, and morphology and were evaluated in a reverse transcription-PCR (RT-PCR) inhibition test. Selected colloids were used for the nonspecific capture of viruses, with RT-PCR being used as a means of detection. Particle size and charge studies allowed us to follow each step of their synthesis, showing the relevance of these methods. The prepared magnetic particles exhibit a good RT-PCR compatibility when used in the range of 5-40 microg in the 50 microl amplification mix. Finally, virus capture tests revealed a capture efficiency of 90% with an average detection sensitivity of 10(3)pfu/ml.

Chromatography, Gel↗

PubMed2005

Monte Carlo assessment of soil moisture effect on high-energy thermal neutron capture gamma-ray by 14N.

Among many conventional techniques, nuclear techniques have shown to be faster, more reliable, and more effective in detecting explosives. In the present work, neutrons from a 5 Ci Am-Be neutron source being in water tank are captured by elements of soil and landmine (TNT), namely (14)N, H, C, and O. The prompt capture gamma-ray spectrum taken by a NaI (Tl) scintillation detector indicates the characteristic photo peaks of the elements in soil and landmine. In the high-energy region of the gamma-ray spectrum, besides 10.829 MeV of (15)N, single escape (SE) and double escape (DE) peaks are unmistakable photo peaks, which make the detection of concealed explosive possible. The soil has the property of moderating neutrons as well as diffusing the thermal neutron flux. Among many elements in soil, silicon is more abundant and (29)Si emits 10.607 MeV prompt capture gamma-ray, which makes 10.829 MeV detection difficult. The Monte Carlo simulation was used to adjust source-target-detector distances and soil moisture content to yield the best result. Therefore, we applied MCNP4C for configuration very close to reality of a hidden landmine in soil.

Journal Article↗

PubMed2005

Sub-micron particle behaviour and capture at an immuno-sensor surface in an ultrasonic standing wave.

The capture of 200 nm biotinylated latex beads from suspensions of concentration 10(7) to 2.5 x 10(8) particle/ml on an immuno-coated surface of the acoustic reflector in an ultrasound standing wave (USW) resonator has been studied while the acoustic pathlength was less than one half wavelength (lambda/2). The particles were delivered to the reflector's surface by acoustically induced flow. The capture dependencies on suspension concentration, duration of experiments and acoustic pressure have been established at 1.09, 1.46 and 1.75 MHz. Five-fold capture increase has been obtained at 1.75 MHz in comparison to the control (no ultrasound) situation. The contrasting behaviours of 1, 0.5 and 0.2 mum fluorescent latex beads in a lambda/4 USW resonator at 1.46 MHz have been characterized. The particle movements were observed with an epi-fluorescent microscope and the velocities of the particles were measured by particle image velocimetry (PIV). The experiments showed that whereas the trajectories of 1 mum particles were mainly affected by the direct radiation force, 0.5 mum particles were influenced both by the radiation force and acoustic streaming. The 0.2 mum latex beads followed acoustic streaming in the chamber and were not detectably affected by the radiation force. The streaming-associated behaviour of the 200 nm particles has implications for enhanced immunocapture of viruses and macromolecules (both of which are also too small to experience significant acoustic radiation force).

Biosensing Techniques↗

PubMed2004

Molecular electron affinities and the calculation of the temperature dependence of the electron-capture detector response.

The use of the electron-capture detector (ECD) to measure molecular electron affinities and kinetic parameters for reactions of thermal electrons is reviewed. The advances of the past decade are emphasized and include the multistate electron-capture detector model and the use of semi-empirical self-consistent field quantum mechanical calculations and half wave reduction potential values to support gas phase experimental results. A procedure for the evaluation of the adiabatic electron affinities of the main group elements and the homonuclear diatomic molecules is presented. Potential excited states are identified for the magnetron (MGN) values for quinones, thermal charge transfer (TCT) values for CS2, C6F6, SF6 and photoelectron spectroscopy (PES) values for O2, NO, nitromethane, and the nucleic acids. Literature electron affinities are then evaluated. The temperature dependence of the electron-capture detector can be calculated using values for kinetic rate constants and electron affinities to optimize response and temperature sensitivity in analytical procedures. The temperature dependence for adenine, guanine, thymine and cytosine are predicted for reactions with thermal electrons. Using the recent advances, the new adiabatic electron affinities are: all in electron volts (eV), 4-F-benzaldehyde (0.57 +/- 0.05) and acetophenones (APs) 4-F-AP (0.52 +/- 0.05); 2-CF3-AP (0.79 +/- 0.05); 3-CF3-AP (0.79 +/- 0.05); 4-CF3-AP (0.89 +/- 0.05); 3-CI-AP (0.67 +/- 0.05); and 4-Cl-AP (0.64 +/- 0.05). The adiabatic electron affinities of chloro and fluorobenzenes range from 0.17 to 1.15 eV and 0.13 to 0.86 eV.

Chemical Phenomena↗

PubMed2004

Hydrophobic interaction chromatography at low salt concentration for the capture of monoclonal antibodies.

We evaluated hydrophobic interaction chromatography (HIC) at low salt concentration for the capture of proteins from feed stocks by using monoclonal antibodies as model samples. It was indicated that the HIC at low salt concentration on critical hydrophobicity supports has a potential for capturing hydrophobic monoclonal antibodies directly from large volumes of feed stocks and recovering bound monoclonal antibodies in high yield. On the other hand, the HIC at low salt concentration did not seem so useful for the capture of weakly hydrophobic monoclonal antibodies. The recovery of weakly hydrophobic monoclonal antibodies from columns packed with critical hydrophobicity supports was not quantitative and significantly decreased as the residence time of the monoclonal antibodies in the columns became longer.

Antibodies, Monoclonal↗

PubMed2005

Aptamer stationary phase for protein capture in affinity capillary chromatography.

The thrombin-binding DNA aptamer was used with thrombin as a model system to investigate protein capture using aptamer stationary phases in affinity capillary chromatography. The aptamer was covalently attached to the inner surface of a bare fused-silica glass capillary to serve as the stationary phase. Proteins were loaded onto the capillary via an applied pressure. The capillary was then washed to remove unbound and non-specifically associated proteins. Finally, the bound protein was released and eluted using 20 mM Tris buffer containing 8 M urea, pH 7.3, at 50 degrees C. Eluate was collected after each step (load, wash and elute) and relative amounts of protein each were compared using fluorescence spectroscopy. The identity of the protein in the collections was confirmed using matrix assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry. The experiment was repeated for thrombin on a bare (unmodified) capillary and a capillary coated with a scrambled-sequence, non-G-quartet forming oligonucleotide that does not bind with thrombin. The results show that the aptamer stationary phase captures approximately three times as much thrombin as the control columns. The experiment was also repeated using human serum albumin (HSA) alone and in an equimolar mixture with thrombin. HSA was not retained on the aptamer capillary, nor did it affect the capture of thrombin from the mixture.

Aptamers, Nucleotide↗