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Biomedical subjects

B E Rice

Publications and source records attributed to B E Rice.

7 recordsLinked to original sources
PubMed2000

A colony lift immunoassay for the specific identification and quantification of Listeria monocytogenes.

A colony lift immunoassay (CLI) has been developed to detect Listeria monocytogenes after the organisms have been cultured on filter membranes or agar plates. Polyvinylidene fluoride membranes (PVDF) (Millipore, Bedford, MA), used in the CLI, were prewet with methanol and used to imprint colonies that were grown on the filter or agar plates. A positive control was applied to the edge of each membrane. The imprinted membranes were subsequently air dried, peroxidase neutralized, blocked, and reacted for 20 min with a 2-microg/ml unconjugated Mab EM-7G1 solution. The membranes were washed briefly and reacted for 30 min with a 1:2000 dilution of a commercially prepared peroxidase-labeled goat anti-mouse secondary antibody (Kirkegaard and Perry Laboratories (KPL), Gaithersburg, MD). After a second wash step, the membranes were exposed to a 3,3',5, 5'-tetramethylbenzidine membrane substrate (KPL), rinsed in deionized water, and allowed to dry. Colonies of L. monocytogenes were identified by a blue color reaction on the membrane, which could be used to reference the colonies either on the filter membranes or agar plates. The CLI was tested against a wide range of Listeria species as well as several non-Listeria species and was shown to have a high degree of sensitivity (96%) and specificity (90%). We have shown that it is useful as a simple and rapid method to detect and identify L. monocytogenes.

Antibodies, Bacterial↗
PubMed2000

Development and evaluation of a 24-hour method for the detection and quantification of Listeria monocytogenes in meat products.

A 24-h filter monitor-based test, Listeria-SELeCT, has been developed to quantify Listeria monocytogenes organisms in meat samples with a sensitivity of < or = 1.0 CFU/g. The technique comprises a filter monitor-based system and a colony lift immunoassay to identify and enumerate the target organism. Meat homogenates were centrifuged and the eluate was filtered to trap and immobilize the microorganisms on the filter. Fraser broth was then added to the filter apparatus to allow the organisms to become established overnight and to inhibit contaminants, after which the filters were transferred onto Modified Oxford medium agar, a selective medium for L. monocytogenes. After 10 to 12 h, a colony lift immunoassay was used to confirm and enumerate suspect colonies on the filter. A correlation study between the Listeria-SELeCT method and the most probable number technique showed the Listeria-SELeCT to be considerably more accurate than the most probable number for quantitatively determining the number of viable organisms in meat samples. Because of ease and speed of testing, the Listeria-SELeCT system also provided major advantages over the most probable number technology.

Animals↗
PubMed1997

Campylobacter jejuni in broiler chickens: colonization and humoral immunity following oral vaccination and experimental infection.

A formalin inactivated, Campylobacter jejuni whole cell vaccine, either with or without Escherichia coli heat labile toxin (LT) as a mucosal adjuvant, was administered orally to broiler chickens. Three vaccine trials were performed, differing in the number of vaccinations, and time of administration, as well as the inclusion and dose of LT. The overall reductions of C. jejuni colonization in the vaccinated chickens ranged from 16 to 93% compared with non-vaccinated controls. Enhanced levels of anti-C. jejuni secretory IgA antibodies were demonstrated in vaccinated chickens. Vaccination also appeared to induce an anamnestic response to C. jejuni antigens in the 14-33 kDa range, as demonstrated by Western immunoblots. Interestingly, the inclusion of LT in the vaccine regimen did not appear to boost the immunogenicity of the vaccine. These results are encouraging and suggest that future development of successful oral vaccines for the control of enteropathogenic Campylobacter in poultry is feasible.

Administration, Oral↗
PubMed1996

Development of a rapid and specific colony-lift immunoassay for detection and enumeration of Campylobacter jejuni, C. coli, and C. lari.

Contamination of retail poultry by Campylobacter spp. is a significant source of human diarrheal disease. We have developed a colony-lift immunoassay (CLI) for the detection of Campylobacter jejuni, C. coli, and C. lari isolated from such sources and grown on selective agar medium or on filter membranes. This technique has been successfully utilized to quantify Campylobacter colonies within 18 to 28 h after sampling. Hydrophobic, high-protein-binding membranes were prewet with methanol and used to imprint bacterial cells from the agar or filter membrane, while leaving colonies intact and viable. The membranes were air dried, peroxidase neutralized, blocked with bovine serum albumin in phosphate-buffered saline, and hybridized for 5 min with an affinity-purified, horseradish peroxidase-labeled goat anti-Campylobacter antibody preparation (Kirkegaard and Perry Laboratories). The membranes were washed briefly, exposed to a 3,'5,5'-tetramethylbenzidine membrane substrate, rinsed in deionized water, and allowed to dry. Lifted colonies of Campylobacter were identified by a blue color reaction on the membrane. Replicas of the membranes were made by marking the location of the Campylobacter colonies on clear transparencies, which were subsequently utilized to locate the original colony on the filter membrane or agar plate. The specificity of this antibody preparation has been evaluated against a wide range of Campylobacter spp., including American Type Culture Collection type and references strains, retail poultry isolates, and isolates obtained from cloacal swabs of live commercial broiler chickens. Specificity against numerous non-Campylobacter spp. obtained from the same sources was also evaluated. The CLI provided a rapid and simple means for detection and enumeration of enteropathogenic Campylobacter organisms. We have successfully combined this CLI procedure with methods recently developed in our laboratories for retail meat and poultry sampling. Potentially, broader applications for use of this technique include detection and enumeration of campylobacters from clinical, veterinary, and environmental samples.

Agar↗