The fluorescent antibody test for the identification of strains of Clostridium botulinum type E

1970 ◽  
Vol 16 (10) ◽  
pp. 917-921 ◽  
Author(s):  
A. C. Emeruwa ◽  
F. E. Ashton ◽  
R. Z. Hawirko
Keyword(s):  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Specific Staining ◽  
Vegetative Cells ◽  
Carbonate Buffer ◽  
Clostridium Botulinum Type E ◽  
Homologous Strain ◽  
The Cross

A fluorescent antibody test for the identification of toxigenic and nontoxigenic type E strains of C. botulinum is described. Hyperimmune rabbit sera were prepared to cell suspensions of the D8, 070, and Beluga strains and to spores of the D8 and PM-15 strains. Fluorescein isothiocyanate immunoglobulin conjugates were prepared and absorbed with C. bifermentans and diluted to eliminate non-specific staining and cross-reactivity with related serotypes. Enzyme pretreatment of spores and washing the stained spores in carbonate buffer, pH 9.0, enhanced the sensitivity of the test. Conjugates of cell antiserum, in most cases, stained the spore bodies and not the vegetative cells of the cross-reacting strains. Conjugates of spore antiserum stained vegetative cells as well as spores but the fluorescence of the cells was eliminated by absorption of the conjugates with young cells of the homologous strain. The absorbed conjugates stained spores of 19/20 isolates from diverse geographical areas without any reduction in the intensity of fluorescence, indicating that spore antigens carry the determinant groups of type E specificity and that the common antigen is a component of the spore body. In addition, some of the cross-reacting strains showed the same degree of fluorescence as the homologous strain, suggesting that more than one antigen may be shared by these strains.

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 An Indirect Fluorescent Antibody Test for the Detection of Antibody to Swine Infertility and Respiratory Syndrome Virus in Swine Sera

1992 ◽  
Vol 4 (2) ◽  
pp. 144-147 ◽  
Author(s):  
In J. Yoon ◽  
Han S. Joo ◽  
William T. Christianson ◽  
Hyun S. Kim ◽  
James E. Collins ◽  
...  
Keyword(s):  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Antibody Test ◽  
Clinical History ◽  
Respiratory Syndrome Virus ◽  
Indirect Fluorescent Antibody ◽  
Swine Farms ◽  
History Of ◽  
Antibody Titers ◽  
Time Of Infection

An indirect fluorescent antibody (IFA) test was developed and standardized to detect and quantitate antibody for swine infertility and respiratory syndrome (SIRS) virus in swine sera. Test results were evaluated using sera of pigs infected both experimentally and naturally with SIRS virus. The IFA test used swine alveolar macrophage (SAM) monolayers prepared in 96-well microplates and infected with SIRS virus. The monolayers were incubated with test sera, washed, and stained with fluorescein isothiocyanate-labeled rabbit anti-swine IgG. After another wash step, the monolayers were examined under a fluorescent microscope. A noninfected SAM control well was included for each sample. The antibody titers for each serum sample were recorded as the highest serum dilutions with specific cytoplasmic fluorescence but no fluorescence in the control wells. To evaluate the test, sera of 4 6-week-old pigs that had been infected with SIRS virus, 2 contact pigs, and 13 experimentally infected sows were used. In the experimentally infected pigs, antibody was first detected at 7 days postexposure (PE) and peaked (1:256–1,024) between 11 and 21 days PE. All 13 sow sera were negative at time of infection but were positive (1:64-> 1: 1,024) at 14–26 days PE. Seven hundred twenty sera collected from 25 different swine farms with or without a history of SIRS were also tested. Of 344 sera from 15 swine farms with a clinical history of SIRS, 257 (74.7%) sera had IFA titers ≥ 1:4, whereas 371 (98.7%) of 376 sera from herds with no history of SIRS were negative. The present results indicate that the IFA is a useful test for the detection and quantitation of SIRS virus antibody in swine sera.

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.

scholarly journals Antigenic Determinants of Alpha-Like Proteins of Streptococcus agalactiae

2004 ◽  
Vol 11 (6) ◽  
pp. 1035-1039 ◽  
Author(s):  
Johan A. Maeland ◽  
Lars Bevanger ◽  
Randi Valsoe Lyng
Keyword(s):  
Western Blotting ◽  
Antigenic Determinant ◽  
Fluorescent Antibody ◽  
Group B Streptococcus ◽  
Antigenic Site ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Antigenic Determinants ◽  
Protective Antibodies ◽  
Group B

ABSTRACT The majority of group B streptococcus (GBS) isolates express one or more of a family of surface-anchored proteins that vary by strain and that form ladder-like patterns on Western blotting due to large repeat units. These proteins, which are important as GBS serotype markers and as inducers of protective antibodies, include the alpha C (Cα) and R4 proteins and the recently described alpha-like protein 2 (Alp2), encoded by alp2, and Alp3, encoded by alp3. In this study, we examined antigenic determinants possessed by Alp2 and Alp3 by testing of antibodies raised in rabbits, mainly by using enzyme-linked immunosorbent assays (ELISA) and an ELISA absorption test. The results showed that Alp2 and Alp3 shared an antigenic determinant, which may be a unique immunological marker of the Alp variants of GBS proteins. Alp2, in addition, possessed an antigenic determinant which showed specificity for Alp2 and a third determinant which showed serological cross-reactivity with Cα. Alp3, in addition to the determinant common to Alp2 and Alp3, harbored an antigenic site which also was present in the R4 protein, whereas no Alp3-specific antigenic site was detected. These ELISA-based results were confirmed by Western blotting and a fluorescent-antibody test. The results are consistent with highly complex antigenic structures of the alpha-like proteins in a fashion which is in agreement with the recently described structural mosaicism of the alp2 and alp3 genes. The results are expected to influence GBS serotyping, immunoprotection studies, and GBS vaccine developments.

scholarly journals Serological cross-reactivity between Anaplasma marginale and an Ehrlichia species in naturally and experimentally infected cattle

2011 ◽  
Vol 23 (6) ◽  
pp. 1181-1188 ◽  
Author(s):  
Batol Al-Adhami ◽  
W. Brad Scandrett ◽  
Vladislav A. Lobanov ◽  
Alvin A. Gajadhar
Keyword(s):  
Fluorescent Antibody ◽  
Surface Protein ◽  
Antibody Test ◽  
Anaplasma Marginale ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Post Inoculation ◽  
Infected Cattle ◽  
Bovine Anaplasmosis ◽  
Major Surface

Seroconversion and cross-reactivity in cattle infected with Anaplasma marginale or a recently described Ehrlichia species (BOV2010 from British Columbia, Canada) were investigated. The study used 76 samples from 20 animals, a commercially available competitive enzyme-linked immunosorbent assay (cELISA) for bovine anaplasmosis, and an indirect fluorescent antibody test (IFAT). Blood smear examination and/or polymerase chain reaction assay were performed to confirm or rule out the presence of Anaplasma or Ehrlichia. Samples comprised 3 groups. Group 1 consisted of 24 samples from 9 cattle naturally infected with Ehrlichia sp. BOV2010. Group 2 had 13 samples from 3 A. marginale–infected cattle from Manitoba, Canada. Group 3 had 39 samples, consisting of 26 from 5 calves experimentally infected with Ehrlichia sp. BOV2010, 10 from 2 calves experimentally infected with A. marginale from cattle (Manitoba) or bison (Saskatchewan), and 3 from an uninfected calf. All samples from cattle naturally or experimentally infected with Ehrlichia sp. BOV2010 or A. marginale were seropositive for A. marginale by both cELISA and IFAT, except 3 calves euthanized at 28 and 33 days post-inoculation (DPI) that did not seroconvert. Antibodies were detected in 2 experimental animals inoculated with Ehrlichia sp. BOV2010, as early as 28 and 33 DPI by the cELISA and IFAT, respectively, and by 42 DPI for both tests. The current study demonstrates that the specificity of the recombinant major surface protein 5 (MSP5) antigen is not restricted to Anaplasma spp., which reduces the utility of the test for serological diagnosis of bovine anaplasmosis in regions where Ehrlichia sp. BOV2010–infected cattle might exist.

scholarly journals STUDIES ON ALKALINE AND ACID RIBONUCLEASES IN MAMMALIAN TISSUES IMMUNOHISTOCHEMICAL LOCALIZATION AND IMMUNOCHEMICAL PROPERTIES

1967 ◽  
Vol 15 (11) ◽  
pp. 662-673 ◽  
Author(s):  
SHIGERU MORIKAWA
Keyword(s):  
Small Intestine ◽  
Fluorescent Antibody ◽  
Fluorescein Isothiocyanate ◽  
Cross Reactivity ◽  
Immunohistochemical Localization ◽  
Rnase A ◽  
Specific Antibodies ◽  
Mammalian Tissues ◽  
Tetramethylrhodamine Isothiocyanate ◽  
Bovine Pancreas

The distribution of alkaline and acid ribonucleases (RNases) was investigated in bovine tissues using fluorescent antibody techniques. The immunochemical properties of the bovine RNases were also examined. Specific antibodies against acid and alkaline RNase obtained by immunizing rabbits with the corresponding antigens were conjugated with fluorescein isothiocyanate or tetramethylrhodamine isothiocyanate. Anti-alkaline RNase antibody reacted against alkaline RNase-A, and anti-acid RNase antibody reacted against two different acid RNases with no cross-reactivity between alkaline and acid RNases. Phosphate-buffered 10% formalin was the best general fixative among those examined, but some differences in staining results were observed depending upon the tissue and fixative employed. Two different patterns of distribution of alkaline RNase in pancreas and kidney were observed depending upon the fixative employed. The localization of RNases in bovine pancreas, liver, small intestine, kidney, spleen, lymph node, thymus, adrenal and heart were investigated and the physiologic roles of the RNases were discussed.

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