Cross-reactivity of environmental bacteria with fluorescent-antibody conjugates forLegionella pneumophila

1984 ◽  
Vol 3 (3) ◽  
pp. 215-215 ◽  
Author(s):  
Y. Glupczynski ◽  
M. Labbe ◽  
E. Yourassowsky
Keyword(s):  
Fluorescent Antibody ◽  
Cross Reactivity ◽  
Antibody Conjugates ◽  
Environmental Bacteria

Fluorescent antibody techniques have allowed for the direct identification and enumeration of individual bacteria in environmental samples without requiring prior growth in culture media (Bahlool and Schmidt 1980, Cloete and Steyn 1988, Macario et al. 1989). The technique involves the use of specific antibodies raised against surface markers of defined pure cultures that are either labelled directly with fluorescent dye molecules or via a fluorescent secondary antibody. This approach has yielded important insights into the spatial distribution of microorganisms, but it suffers from a number of disadvantages. For example, expression of the antigen may be influenced by environmental factors; false-positive and false-negative results may be obtained due to cross-reactivity or lack of reaction; non-specific binding of antibodies may result in high levels of background fluorescence; and production of specific antibodies requires a pure culture of the organism of interest (Cloete and de Bruyn Various recombinant DNA techniques have subsequently been developed that are independent of cultivation methods (Fig. 1). These techniques provide ways of detecting and quantifying specific phylogenetic groups of microbes on 16S rDNA sequences, and relevant structural genes provide ways of monitoring microbial populations of environmental and industrial systems. In addition to these tools, a number of emerging technologies such as the use of biomarker genes are being increasingly used to monitor with great precision and accuracy the behaviour of microbes in the environment.

2003 ◽  
pp. 218-219
Keyword(s):  
Recombinant Dna ◽  
Culture Media ◽  
Specific Binding ◽  
Fluorescent Antibody ◽  
False Negative ◽  
Cross Reactivity ◽  
Phylogenetic Groups ◽  
Specific Antibodies ◽  
Negative Results ◽  
Recombinant Dna Techniques

scholarly journals Rapid Fluorescein and Protein Assay Method for Fluorescent-Antibody Conjugates

1966 ◽  
Vol 14 (2) ◽  
pp. 271-275
Author(s):  
Arthur F. Wells ◽  
Curtis E. Miller ◽  
Marvin K. Nadel
Keyword(s):  
Fluorescent Antibody ◽  
Assay Method ◽  
Protein Assay ◽  
Antibody Conjugates

scholarly journals IMMUNOHISTOCHEMICAL LOCALIZATION OF CATALASE IN MAMMALIAN TISSUES

1969 ◽  
Vol 17 (1) ◽  
pp. 30-35 ◽  
Author(s):  
SHIGERU MORIKAWA ◽  
TAKAYUKI HARADA
Keyword(s):  
Fluorescent Antibody ◽  
Bovine Liver ◽  
Cross Reactivity ◽  
Immunohistochemical Localization ◽  
Hepatic Cells ◽  
Surface Active Agents ◽  
Erythrocyte Catalase ◽  
Mammalian Tissues ◽  
Splenic Cells ◽  
Proximal Tubuli

The distribution of catalase was investigated in bovine tissues using fluorescent antibody techniques. The immunochemical properties of liver catalase were also examined. Two distinct components of liver catalase in immune system were found. Both of them possessed common antigenicity with erythrocyte catalase. No cross-reactivity was observed by immunodiffusion between catalase and the other hemoproteins such as lactoperoxidase, cytochrome c and hemoglobins. Bovine liver, pancreas, kidney, spleen and peripheral blood were examined. Catalase was located mainly in the cytoplasm of hepatic cells, acinar cells of the pancreas, epithelia of proximal tubuli, splenic cells scattered in the red pulp and some leukocytes. It was not found in any nucleus. Intracorpuscular catalase could be revealed in the erythrocytes treated with surface-active agents but not in frozen sections.

scholarly journals Evaluation of Multivalent Leptospira Fluorescent Antibody Conjugates for General Diagnostic Use

1989 ◽  
Vol 1 (2) ◽  
pp. 146-149 ◽  
Author(s):  
David A. Miller ◽  
Mark A. Wilson ◽  
Clyde A. Kirkbride
Keyword(s):  
Fluorescent Antibody ◽  
Bacterial Species ◽  
Antibody Test ◽  
Additional Method ◽  
Diagnostic Laboratory ◽  
Bacterial Cultures ◽  
Typical Cell ◽  
Antibody Conjugates ◽  
Treponema Hyodysenteriae ◽  
Culture Positive

Four lots of conjugate were evaluated for optimal dilution and degree of fluorescence produced with reference cultures and bovine and porcine leptospira isolates. One lot that uniformly produced better fluorescence was evaluated for sensitivity and specificity with reference cultures, isolates, culture-positive tissues, and 13 other bacterial species. Further evaluation of the conjugates was done with bovine, porcine, and ovine specimens submitted to a diagnostic laboratory. Leptospires were detected with the fluorescent antibody test (FAT) in 9 of 21 culture-positive bovine kidneys and were detected in diluted cultures when present at concentrations of 102-1O3 organisms/ml. With the exception of Treponema hyodysenteriae, FAT's of other bacterial cultures produced minimal fluorescence or were negative. Positives were characterized by moderate to brilliant fluorescence of typical cell forms, and most nonspecific fluorescence was eliminated with a flazo-orange counter-stain. The results indicated that the FAT utilizing multivalent conjugates could be used successfully as an additional method for diagnosis of leptospira infections.

scholarly journals Impact of antigenic diversity on laboratory diagnosis of Avian bornavirus infections in birds

2014 ◽  
Vol 26 (6) ◽  
pp. 769-777 ◽  
Author(s):  
Vanessa Zimmermann ◽  
Monika Rinder ◽  
Bernd Kaspers ◽  
Peter Staeheli ◽  
Dennis Rubbenstroth
Keyword(s):  
Matrix Protein ◽  
Fluorescent Antibody ◽  
Laboratory Diagnosis ◽  
Cross Reactivity ◽  
Diagnostic Tools ◽  
Phylogenetic Groups ◽  
Antigenic Diversity ◽  
Antibody Assays ◽  
Causative Agents ◽  
Antibody Titers

Avian bornaviruses (ABVs) are a group of genetically diverse viruses within the Bornaviridae family that can infect numerous avian species and represent the causative agents of proventricular dilatation disease, an often fatal disease that is widely distributed in captive populations of parrots and related species. The current study was designed to assess the antigenic variability of the family Bornaviridae and to determine its impact on ABV diagnosis by employing fluorescent antibody assays. It was shown that polyclonal rabbit sera directed against recombinant bornavirus nucleoprotein, X protein, phosphoprotein, and matrix protein provided sufficient cross-reactivity for the detection of viral antigen from a broad range of bornavirus genotypes grown in cell culture. In contrast, a rabbit anti-glycoprotein serum and 2 monoclonal antibodies directed against nucleoprotein and phosphoprotein proteins reacted more specifically. Antibodies were readily detected in sera from avian patients infected with known ABV genotypes if cells persistently infected with a variety of different bornavirus genotypes were used for analysis. For all sera, calculated antibody titers were highest when the homologous or a closely related target virus was used for the assay. Cross-reactivity with more distantly related genotypes of other phylogenetic groups was usually reduced, resulting in titer reduction of up to 3 log units. The presented results contribute to a better understanding of the antigenic diversity of family Bornaviridae and further emphasize the importance of choosing appropriate diagnostic tools for sensitive detection of ABV infections.

scholarly journals Indirect hemagglutination test for human antibody to typhus and spotted fever group rickettsiae

1975 ◽  
Vol 2 (5) ◽  
pp. 430-437
Author(s):  
A Shirai ◽  
J W Dietel ◽  
J V Osterman
Keyword(s):  
Complement Fixation ◽  
Spotted Fever ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Human Antibody ◽  
Spotted Fever Group ◽  
Rickettsia Rickettsii ◽  
Indirect Hemagglutination ◽  
Indirect Hemagglutination Test

An indirect hemagglutination (IHA) test is described that uses glutaraldehyde-stabilized erythrocytes treated with a rickettsial erythrocyte-sensitizing substance obtained from Rickettsia typhi or Rickettsia rickettsii. The serological reagent was stable for at least 3 months at room temperature and 6 months at 4 C. It exhibited group specificity and no group cross-reactivity. At a minimum dilution of 1:40, acute and early convalescent epidemic and murine typhus antisera showed 86% positive reactors, whereas similar spotted fever antisera had 74% positive reactors. In comparison with the indirect fluorescent antibody test, the IHA procedure gave lower titers but showed comparable detection of seroconversion with most paired sera. The IHA test demonstrated significantly higher titers than the complement fixation test and was more sensitive than either the complement fixation or Weil-Felix test in identifying seroconversion. No agglutination was observed when sensitized erythrocytes were tested with rodent sera known to contain rickettsial antibodies.

scholarly journals Preparation of Agglutinating Antisera and Fluorescent-Antibody Conjugates Against Pasteurella tularensis in Equines

1970 ◽  
Vol 19 (6) ◽  
pp. 894-897
Author(s):  
James H. Green ◽  
Richard C. Bolin ◽  
Russell K. Carver ◽  
Herman Gross ◽  
Nan Pigott ◽  
...  
Keyword(s):  
Fluorescent Antibody ◽  
Antibody Conjugates

scholarly journals Novel Indirect Fluorescent Antibody Test for Lyme Disease

1996 ◽  
Vol 8 (2) ◽  
pp. 196-201 ◽  
Author(s):  
Margaret A. Chambers ◽  
Larry J. Swango ◽  
James C. Wright
Keyword(s):  
Human Error ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Fluorescent Antibody ◽  
Serum Samples ◽  
Specific Pathogen ◽  
Membrane Bound ◽  
Animal Use

An indirect fluorescent antibody (IFA) test was developed using a novel format of Borrelia burgdorferi organisms adhered to a monolayer of cultured endothelial cells derived from an equine tumor. Sensitivity and specificity of the new IFA test for detecting anti- B. burgdorferi antibodies were evaluated using sera from dogs inoculated with live B. burgdorferi or vaccinated with B. burgdorferi bacterin or leptobacterins and from unvaccinated specific-pathogen-free (SPF) dogs. To compare the new IFA test with existing tests, serum samples were submitted to independent laboratories to be tested by enzyme-linked immunosorbent assay (ELISA) and a traditional IFA test. Samples were also tested with 2 commercially available membrane-bound ELISA kits. Both Borrelia-inoculated dogs and dogs vaccinated with B. burgdorferi bacterin developed levels of antibody detectable by the new IFA test. Dogs vaccinated with a combination canine vaccine or leptobacterin for food animal use developed detectable levels of antibody against Leptospira but remained seronegative for Borrelia by the new IFA test, as did the unvaccinated SPF dogs. The new IFA test was sensitive, detecting antibodies against B. burgdorferi as early as 7 days postinoculation. It was also specific, showing no cross-reactivity with anti- Leptospira antibodies induced by vaccination with leptobacterins. The new IFA test compared favorably with both the standardized traditional IFA test and ELISA. Results from both membrane-bound ELISA kits were not consistent when compared with each other or with the new IFA test. The new IFA test had low nonspecific fluorescence, which made it easier to evaluate and reduced the human error and variability of test results.

Myxosoma cerebralis: Fluorescent Antibody Techniques for Antigen Recognition

1978 ◽  
Vol 35 (6) ◽  
pp. 828-832 ◽  
Author(s):  
Maria E. Markiw ◽  
Ken Wolf
Keyword(s):  
Life Cycle ◽  
Key Words ◽  
Fluorescent Antibody ◽  
Cross Reactivity ◽  
Antigen Recognition ◽  
Fluorescent Antibody Technique ◽  
Indirect Fluorescent Antibody ◽  
Whirling Disease ◽  
Antibody Technique ◽  
Direct Fluorescent Antibody

Rabbits were immunized with antigens extracted from mature spores or prespore stages of Myxosoma cerebralis, and the resulting antisera and their globulins were used in direct and indirect fluorescent antibody techniques. Both kinds of antisera reacted with homologous spores and with stages of the organism that precede spores. When tested for specificity against spores of 12 other myxosporidans the direct fluorescent antibody technique showed cross-reactivity with only one other and that was a species of Myxosoma. The indirect fluorescent antibody technique showed some reactions across generic lines. The antisera have application in studies of the parasite's life cycle and in diagnostics. Key words: spores, parasites, direct fluorescent antibody techniques, indirect fluorescent antibody techniques, diagnosis, myxosporidans, whirling disease

Sign in / Sign up

Export Citation Format

Share Document