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Biomedical subjects

C A Rowe

Publications and source records attributed to C A Rowe.

11 recordsLinked to original sources
PubMed1999

Array biosensor for simultaneous identification of bacterial, viral, and protein analytes.

The array biosensor was fabricated to analyze multiple samples simultaneously for multiple analytes. The sensor utilized a standard sandwich immunoassay format: Antigen-specific "capture" antibodies were immobilized in a patterned array on the surface of a planar waveguide and bound analyte was subsequently detected using fluorescent tracer antibodies. This study describes the analysis of 126 blind samples for the presence of three distinct classes of analytes. To address potential complications arising from using a mixture of tracer antibodies in the multianalyte assay, three single-analyte assays were run in parallel with a multianalyte assay. Mixtures of analytes were also assayed to demonstrate the sensor's ability to detect more than a single species at a time. The array sensor was capable of detecting viral, bacterial, and protein analytes using a facile 14-min assay with sensitivity levels approaching those of standard ELISA methods. Limits of detection for Bacillus globigii, MS2 bacteriophage, and staphylococcal enterotoxin B (SEB) were 10(5) cfu/mL, 10(7) pfu/mL, and 10 ng/mL, respectively. The array biosensor also analyzed multiple samples simultaneously and detected mixtures of the different types of analytes in the multianalyte format.

Antibodies↗
PubMed1999

An array immunosensor for simultaneous detection of clinical analytes.

A fluorescence-based immunosensor has been developed for simultaneous analysis of multiple samples. A patterned array of recognition elements immobilized on the surface of a planar waveguide is used to "capture" analyte present in samples; bound analyte is then quantified by means of fluorescent detector molecules. Upon excitation of the fluorescent label by a small diode laser, a CCD camera detects the pattern of fluorescent antigen:antibody complexes on the sensor surface. Image analysis software correlates the position of fluorescent signals with the identity of the analyte. This immunosensor was used to detect physiologically relevant concentrations of staphylococcal enterotoxin B (SEB), F1 antigen from Yersinia pestis, and D-dimer, a marker of sepsis and thrombotic disorders, in spiked clinical samples.

Antigens, Bacterial↗
PubMed1999

Array biosensor: optical and fluidics systems.

Optical and fluidics systems have been developed as central components for an automated array biosensor. Disposable planar waveguides are patterned with immobilized capture antibodies using a physically isolated patterning (PIP) method. The PIP method enables simultaneous deposition of several antibodies and completely circumvents cross-immobilization problems encountered with other array deposition processes. A multi-channel fluidics cell allows numerous assays to be performed on the patterned waveguide. The sensing arrays are optically interrogated using a diode laser with a tailored output to optimize coupling to and maximize excitation uniformity within the waveguide. A patterned cladding is employed to optically isolate the waveguide from perturbations induced by the permanently attached flow cells. Compact optics image the evanescently excited fluorescence onto a large area, cooled CCD array. The image data is processed and automated signal analysis corrects for local background and noise variations.

Journal Article↗
PubMed1998

Rapid detection of D-dimer using a fiber optic biosensor.

We describe a rapid and sensitive method for detection and quantification of D-dimer and other crosslinked fibrin degradation products (XL-FDPs), which are present in elevated concentrations in patients with sepsis and thrombotic disorders. The method utilizes a sandwich fluoroimmunoassay immobilized in the sensing region of an evanescent wave biosensor. Physiological concentrations of D-dimer and high molecular weight XL-FDP could be determined in buffer and plasma samples on calibrated fibers in 11 min. Samples from septic patients were assayed using ELISA and the fiber optic method; concentrations determined by fiber optic assay were strongly correlated with those determined by ELISA (r = 0.918); intra- and inter-assay errors were comparable to those from ELISAs. Given its accuracy and rapid response time, this fiber optic biosensor shows great potential for development as a diagnostic tool.

Biosensing Techniques↗
PubMed1996

Oral immunization with a replication-deficient recombinant vaccinia virus protects mice against influenza.

Mice immunized with two intragastrically administered doses of a replication-deficient recombinant vaccinia virus containing the hemagglutinin and nucleoprotein genes from H1N1 influenza virus developed serum anti-H1 immunoglobulin G (IgG) antibody that completely protected the lungs from challenge with H1N1. Almost all of the mice given two intragastric doses also developed mucosal anti-H1 IgA antibody, and those with high anti-H1 IgA titers had completely protected noses. Intramuscular injection of the vaccine protected the lungs but not the noses from challenge. We also found that the vaccine enhanced recovery from infection caused by a shifted (H3N2) influenza virus, probably through the induction of nucleoprotein-specific cytotoxic T-lymphocyte activity. A replication-deficient, orally administered, enteric-coated, vaccinia virus-vectored vaccine might safely protect humans against influenza.

Administration, Oral↗
PubMed1993

Characterization of monoclonal antibodies against a type SAT 2 foot-and-mouth disease virus.

This paper is the first to describe characterization of monoclonal antibodies (MAbs) against a South African Territories 2 (SAT 2) foot-and-mouth disease virus (isolate Rho 1/48). Twelve MAbs which neutralized homologous virus were characterized in indirect and sandwich ELISA using purified Rho 1/48 virus particles, subunits, trypsin-treated, and chemically denatured virus. All the MAbs inhibited haemagglutination by parental virus. Binding of the MAbs to 73 SAT 2 field isolates was measured in a sandwich ELISA and defined four distinct antigenic regions. Preliminary characterization of escape mutants selected with some of the MAbs using virus neutralization tests, ELISA, and amino acid sequencing is included. MAbs 2, 25, 40, 48 and 64, reacted with a linear epitope on the VP1 loop region. An amino acid change at position 149 (valine to glutamic acid) was detected in mutants selected by MAb 2 and 40 and this eliminated binding and neutralization by all the other MAb. This epitope was conformation-dependent and was conserved in all 73 isolates of SAT 2 examined. Escape mutants isolated with MAb 41 and 44, had changes at positions 156 (glycine to aspartic acid), or 158 (serine to leucine) respectively. These MAbs bound with Rho 1/48 only out of 73 field strain viruses studies and the reactions of MAbs from the other groups was unaltered. MAb 27, 28 and 37 reacted with a conformation-dependent epitope on VP1 which was not conserved in field isolates. All mutants selected by these MAbs had a single amino acid substitution at position 149 (valine to alanine). The same change was always found in field isolates which did not bind MAbs from this group. MAb 11 reacted with a linear epitope associated with amino acids 147 or 148 on VP1 and showed similar binding characteristics to a conformation dependent MAb 7, no amino acid residue changes were found within VP1 for monoclonal antibody 7 mutants.

Amino Acid Sequence↗
PubMed1991

Non-pathogenic strains of Aujeszky's disease virus are not reactivated in swine after corticosteroid treatment.

1. The Bartha-K and NIA-4 strains of Aujeszky's disease virus (ADV) were readily isolated from oropharyngeal swabs up to 7 days after intranasal vaccination of young piglets. 2. Neither strain could be reisolated 14 days after starting treatment with 10 mg of the corticosteroid isoflupredone acetate per kg of body weight, administered intramuscularly for 4 consecutive days when pigs were 7-9 months of age. 3. Similar treatment with corticosteroid pigs infected with two virulent ADV strains resulted in the reactivation of infection and recovery of ADV from oropharyngeal swabs. 4. Serum neutralizing antibodies were present in all piglets vaccinated twice (2 week interval) intranasally with the attenuated ADV strains, 4 weeks after primary vaccination. However, these antibodies were no longer detectable in some pigs at 12 (NIA-4) and 20 (Bartha-K) weeks of age even in undiluted sera. 5. Neutralizing antibodies resulting from infection with virulent ADV were always detectable, were higher in titer than those produced by the vaccine strains and did not vary in a clear pattern after corticosteroid treatment. 6. These results indicate that the Bartha-K and NIA-4 strains undergo little or no latency in swine and confirm the latency of virulent strains of ADV.

Adrenal Cortex Hormones↗
PubMed1989

Experimental vaccination of rabbits with non-pathogenic strains of Aujeszky's disease virus.

1. The Bartha K and NIA-4 strains of Aujeszky's disease virus (ADV) were non-pathogenic for rabbits vaccinated once or twice by nasal instillation or intramuscular injection. Neutralizing antibodies were detected in 68% of the rabbits two weeks after primary vaccination and in all rabbits at challenge. 2. Challenge doses of virulent ADV greater than 10(5.0) median tissue culture infective doses (TCID50) resulted in the death of most vaccinated and all unvaccinated rabbits with typical signs of Aujeszky's disease within 4 days. ADV was recovered from brain and lung suspensions of vaccinated and unvaccinated rabbits who had died as a result of the challenge. 3. When the challenge dose was reduced to approximately 10(3.0) TCID50, rabbits vaccinated twice survived while all unvaccinated controls died within 3 days.

Animals↗
PubMed1983

Transmission of bovine leukaemia virus in milk.

Nineteen calves born to dams free of bovine leukaemia virus (BLV) did not possess maternally derived precipitating antibody to BLV in their sera after the ingestion of colostrum. Eight of these calves remained serologically negative after being fed milk from BLV-free cows while three (27.3%) of 11 similar calves that had been fed milk from BLV-infected cows developed antibody. Forty-four of 47 calves born to BLV-infected dams acquired maternal antibody to BLV after ingesting colostrum. Two (8.7%) of the 23 calves fed milk from BLV-free cows developed antibody to BLV probably as a result of transplacental or colostrum infection whereas four (16.7%) of the 24 calves fed milk from BLV-infected cows developed antibody. It is concluded that milk transmission of BLV is responsible in part for the high rates of infection encountered in our dairy herds and that calves lacking specific maternal antibody are more susceptible to BLV infection through the ingestion of milk than are calves with maternal antibody.

Animals↗
PubMed1981

Enzootic bovine leukosis virus in Brazil.

A sero-epidemiological survey for antibodies to the glycoprotein of enzootic bovine leukosis virus showed that the infection is widely disseminated in the State of Rio de Janeiro, Brazil. Sero from 1,290 females and 154 males from 12 dairy herds were tested by the agar gel precipitin test. Seven hundred and one females (54.3%) and 68 males (44.2%) were found to have specific antibodies. These antibodies were demonstrated in all 7 age groups tested. The older age groups contained the highest percentage of reactors. The results are briefly discussed in relation to management practices and environmental conditions.

Aging↗