scholarly journals Characterization of Serological Responses to Pertussis

2006 ◽  
Vol 13 (3) ◽  
pp. 341-348 ◽  
Author(s):  
Mineo Watanabe ◽  
Beverly Connelly ◽  
Alison A. Weiss
Keyword(s):  
Adenylate Cyclase ◽  
Serological Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Vaccine Antigens ◽  
History Of ◽  
Antibody Titers ◽  
Iga Antibody ◽  
Conventional Vaccine ◽  
Culture Positive

ABSTRACT We have compared the use of five nonvaccine antigens to the use of conventional vaccine antigens, pertussis toxin (PT), and filamentous hemagglutinin (FHA) for the serological diagnosis of pertussis by enzyme-linked immunosorbent assay (ELISA). The nonvaccine antigens included the catalytic region of adenylate cyclase toxin (CatACT), the C-terminal region of FHA (C-FHA), lipooligosaccharide (LOS), the peptidoglycan-associated lipoprotein (PAL), and the BrkA protein. The serological responses of individuals with culture-confirmed pertussis were compared to those of adults with no recent history of a coughing disease. An immunoglobulin G (IgG) ELISA for PT was the most sensitive (92.2%) test for the serodiagnosis of pertussis. Of the nonvaccine antigens, ELISA for IgG responses to CatACT (sensitivity, 62.8%), C-FHA (sensitivity, 39.2%), and LOS IgA (sensitivity, 29.4%) were less sensitive but could also distinguish culture-positive individuals from control individuals. The use of a combination of multiple ELISA targets improved the sensitivity of the assay for serological diagnosis. Elevated IgG and IgA antibody titers persisted for more than a year in the individuals with culture-confirmed pertussis.

Study of Antibody Levels in Children with Purulent Otitis Media

1980 ◽  
Vol 89 (3_suppl) ◽  
pp. 117-120 ◽  
Author(s):  
P. Branefors ◽  
T. Dahlberg ◽  
O. Nylén
Keyword(s):  
Otitis Media ◽  
Serum Antibody ◽  
Acute Otitis Media ◽  
High Serum ◽  
Haemophilus Influenzae Type ◽  
Enzyme Linked Immunosorbent Assay ◽  
Polysaccharide Antigens ◽  
Antibody Titers ◽  
Iga Antibody ◽  
Antibody Levels

A series of episodes of acute otitis media were studied with reference to the bacterial findings in the nasopharynx and the specific antibody response in a group of children nine months to ten years of age, with previous frequent episodes of acute otitis media, Serum IgG, IgM and IgA antibody levels against five polysaccharide antigens, namely Haemophilus influenzae type b and Streptococcus pneumoniae types 3, 6, 19 and 23, were studied by means of an enzyme-linked immunosorbent assay. The selection of polysaccharide antigens was based on isolation frequency. The sera to be tested were tenfold serially diluted. An extinction of 0.2 over the base was taken as the end-point titer and expressed as in-log10. The results showed that most children including those under three years of age showed increasing homologous antibody titers at an infection, or had already initially very high antibody titers, especially of the IgG class. The titers reached levels of 104 to 105. In some cases, however, it could be shown that high serum antibody titers did not give protection against a new infection with the same serological type of bacteria. It was also demonstrated that most children, regardless of age, had IgG and IgM titers against the heterologous antigens. In some cases the levels were quite high (103 to 104). However, the IgA antibody levels were lower and in a considerable number of samples antibodies were not even detectable.

Production and Characterization of Antibodies against Fumonisin B1

1996 ◽  
Vol 59 (9) ◽  
pp. 992-997 ◽  
Author(s):  
FENG-YIR YU ◽  
FUN S. CHU
Keyword(s):  
Detection Limit ◽  
Polyclonal Antibodies ◽  
Fumonisin B1 ◽  
Correlation Coefficients ◽  
Keyhole Limpet Hemocyanin ◽  
Hplc Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Competitive Elisa ◽  
Antibody Titers

Polyclonal antibodies against fumonisin B1 (FmB1) were produced in rabbits after immunizing the animals with either FmBl-keyhole limpet hemocyanin (KLH) or FmB1 bovine serum albumin (BSA). A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) and an indirect competitive ELISA (idc-ELISA) were used for the characterization of the antibodies and for analysis of the toxin in corn samples. The antibody titers in the serum of rabbits immunized with FmBl-KLH were considerably higher than in those immunized with FmBl-BSA. The antibodies from the rabbits immunized with FmBl-KLH were further characterized. The concentrations causing 50% inhibition of binding of FmB1-horseradish peroxidase (HRP) to the antibodies by FmB1, FmB2 and FmB3 in the ELISA were found to be 0.45, 0.72, and 25 ng/ml, respectively. The detection limit of FmBl, based on 95% confidence at 5% of inhibition of binding of FmBl-HRP conjugate, in buffer of the dc-ELISA was found to be 0.05 ng/ml. In the presence of a matrix such as corn, the detection limit was less than 50 ppb. The overall analytical recoveries of FmBl (50 to 1,000 ng/g) added to the ground corn and then extracted with CH3CN/H2O (1/1, vol/vol) with cleanup and without cleanup in the dc ELISA were found to be 70.5 and 85.9%, respectively. A good correlation was found between the FmBl levels in 2 starch and 10 naturally contaminated corn samples analyzed by the dc-ELISA and the high-pressure liquid chromatography (HPLC) method. The correlation coefficients between ELISA and HPLC were found to be 0.955 (y [ELISA] = 1.3 1x [HPLC] + 77 ppb; P < 0.001) and 0.811 (y = 1.13x + 34 ppb; P < 0.01) for the sample without and with cleanup treatment, respectively.

Production and Characterization of Antibodies Against Neosaxitoxin

1992 ◽  
Vol 75 (2) ◽  
pp. 341-345 ◽  
Author(s):  
Fun S Chu ◽  
Xuan Huang ◽  
Sherwood Hall
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Antibody ◽  
Antibody Titers ◽  
Antibody Elisa

Abstract Antibody against neosaxitoxin (neo-STX) was obtained from rabbits after immunization with neo- STX conjugated to either keyhole limpet hemocyanln (KLH) or bovine serum albumin (BSA). An indirect enzyme-linked Immunosorbent assay (ELISA), In which either neo-STX-BSA or neo-STXKLH was coated to the mlcroplate, was used to monitor the antibody titer. Although high antibody titers were obtained from rabbits after immunization with both immunogens, only antibody obtained from rabbits immunized with neo-STX-KLH was useful for Immunoassay. Competitive indirect ELISA revealed that the antibodies obtained from rabbits Immunized with neo-STX-KLH are specific for neo-STX but also have good cross-reactivity with STX. The concentrations causing 50% inhibition binding of neo-STX-BSA to the anti-neo-KLH by neo-STX, STX, and decarbamoyl-STX (DC-STX) were 0.9,8.0, and 53.1 ng/mL, respectively. Saxitoxin conjugated to polylyslne (STX-PLL) was also used as the coating reagent In the indirect ELISA. The concentrations causing 50% inhibition binding of antl-neo-STX-KLH to STX-PLL coated on the mlcrotiter plate by neo-STX, STX, and DC-STX were 1.2,4.1, and 36.1 ng/mL, respectively. With this newly developed antibody, ELISA could be a very effective method for monitoring seafood for both neo-STX and STX.

scholarly journals Persistence ofLeishmania donovaniAntibodies in Past Visceral Leishmaniasis Cases in India

2010 ◽  
Vol 18 (2) ◽  
pp. 346-348 ◽  
Author(s):  
Kamlesh Gidwani ◽  
Albert Picado ◽  
Bart Ostyn ◽  
Shri Prakash Singh ◽  
Rajiv Kumar ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Immunosorbent Assay ◽  
Leishmania Donovani ◽  
Extended Period ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Agglutination Test ◽  
History Of ◽  
Antibody Titers

ABSTRACTThe persistence of anti-Leishmania donovaniantibodies in past visceral leishmaniasis (VL) cases was retrospectively assessed by means of the direct agglutination test (DAT) and the rK39 enzyme-linked immunosorbent assay (ELISA). Antibody titers remained high for an extended period of time in past cases of VL. These results highlight the need to carefully elicit the history of patients with VL symptoms.

scholarly journals Characterization of New Formalin-Detoxified Botulinum Neurotoxin Toxoids

2008 ◽  
Vol 15 (9) ◽  
pp. 1374-1379 ◽  
Author(s):  
James E. Keller
Keyword(s):  
Botulinum Neurotoxin ◽  
Lethal Dose ◽  
Enzyme Linked Immunosorbent Assay ◽  
Reaction Conditions ◽  
Inhibition Elisa ◽  
Naturally Occurring ◽  
Antibody Titers

ABSTRACT Antigenicities of several formalin-detoxified botulinum neurotoxin preparations were measured by inhibition and sandwich enzyme-linked immunosorbent assay (ELISA), and immunogenicity was studied in mice. The toxoids were derived primarily from the serotype A 150-kDa neurotoxin protein, while one toxoid was derived from the naturally occurring 900-kDa toxin-hemagglutinin complex. Antigenicity was severely compromised in two commercially available toxoids. A variety of new toxoids were synthesized in-house by optimizing formaldehyde reaction conditions. Three of the resulting toxoids were found to be antigenically identical to the native toxin, as measured by inhibition ELISA, in spite of showing a reduction of toxicity by more than 100,000-fold. Sandwich ELISAs indicated that the in-house toxoids were two- to threefold less antigenic than the neurotoxin compared to commercial toxoids, which were about 100-fold less antigenic. Mice were immunized twice, on day 0 and day 14. By day 28, relatively high toxin-specific immunoglobulin G (IgG) titers were detected in animals that had received any of the in-house toxoids, with greater than 99% being IgG1 and the remainder being IgG2. These immunized mice remained asymptomatic after being challenged with 50 to 1,000,000 50% lethal dose (LD50) units of the 900-kDa neurotoxin. In contrast, animals immunized with several different batches of commercially available toxoids did not develop measurable toxin-specific antibody titers. However, these mice survived neurotoxin challenges with 2 LD50 units but died when challenged with 6 LD50 units. Neutralizing titers measured from pools of sera generated with the in-house toxoid preparations ranged from 2.5 to 5 U/ml. In terms of predicting immunogenicity, inhibition ELISAs comparing each formalin toxoid to the parent toxin provided good insight for screening the new toxoids as well as for estimating their relative in vivo potencies. Inhibition ELISA data indicate that those toxoids that most closely resemble the native toxin are highly immunogenic and protective. The superior quality of these new toxoids makes them useful tools for continued use in ELISA development and for antitoxin production.

scholarly journals A Sensitive Immunoblotting Technique for the Serodiagnosis of Brucella Ovis Infections

1994 ◽  
Vol 6 (2) ◽  
pp. 188-194 ◽  
Author(s):  
Reinhold Kittelberger ◽  
Mike Hansen ◽  
Gail P. Ross ◽  
Frans Hilbink
Keyword(s):  
Sodium Dodecyl ◽  
Enzyme Linked Immunosorbent Assay ◽  
Brucella Ovis ◽  
Specific Pathogen ◽  
History Of ◽  
Molecular Masses ◽  
Culture Positive ◽  
Gel Diffusion ◽  
Disease Free ◽  
Immunoblotting Technique

A simplified electrophoretic immunoblotting technique based on antigen extracted from Brucella ovis cells with sodium dodecyl sulfate/mercaptoethanol was compared with the complement fixation test (CFT), the enzyme-linked immunosorbent assay, and the gel diffusion test. Sera from 89 chronically infected, semen culture-positive rams, 378 sera from B. ovis-infected flocks, 300 sera from accredited disease-free flocks, and 29 sera from specific-pathogen-free sheep were used. The immunoblotting technique had sensitivity and specificity comparable to those of the standard tests and was able to identify several CFT-negative or -borderline sera as positive. The major immunoreactive antigens of B. ovis had molecular masses of 63,29, 19 kD (proteins) and 8-12 kD (rough lipopolysaccharide). Antibodies against these antigens were present in 96% of CFT-positive sera from infected flocks and in 100% of sera from semen culture-positive rams. However, immunoblotting also identified antibodies to components other than the major antigens in 1% of CFT-negative sera from infected flocks and in 7.7% of the sera from flocks with a history of freedom from the disease. These reactions probably represent cross-reactivities with other microorganisms and were distinguishable from truly positive reactions.

TBE in Sweden

2020 ◽  
Keyword(s):  
Genetic Characterization ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Second Phase ◽  
Serum Samples ◽  
Tick Borne Encephalitis ◽  
Specific Immunoglobulin ◽  
Tick Borne Encephalitis Virus ◽  
First Time

Tick-borne encephalitis virus (TBEV) was isolated for the first time in Sweden in 1958 (from ticks and from 1 tick-borne encephalitis [TBE] patient).1 In 2003, Haglund and colleagues reported the isolation and antigenic and genetic characterization of 14 TBEV strains from Swedish patients (samples collected 1991–1994).2 The first serum sample, from which TBEV was isolated, was obtained 2–10 days after onset of disease and found to be negative for anti-TBEV immunoglobulin M (IgM) by enzyme-linked immunosorbent assay (ELISA), whereas TBEV-specific IgM (and TBEV-specific immunoglobulin G/cerebrospinal fluid [IgG/CSF] activity) was demonstrated in later serum samples taken during the second phase of the disease.

Subversive Mobilities

Transfers ◽  
2013 ◽  
Vol 3 (1) ◽  
pp. 7-25 ◽  
Author(s):  
Mikkel Thelle
Keyword(s):  
Public Space ◽  
Cultural History ◽  
Turn Of The Century ◽  
Public Action ◽  
History Of ◽  
Urban Conflict

The article approaches mobility through a cultural history of urban conflict. Using a case of “The Copenhagen Trouble,“ a series of riots in the Danish capital around 1900, a space of subversive mobilities is delineated. These turn-of-the-century riots points to a new pattern of mobile gathering, the swarm; to a new aspect of public action, the staging; and to new ways of configuring public space. These different components indicate an urban assemblage of subversion, and a new characterization of the “throwntogetherness“ of the modern public.

Shakespeare Afoot

2018 ◽  
Author(s):  
John Kerrigan
Keyword(s):  
Early Modern ◽  
Richard Iii ◽  
Midsummer Night's Dream ◽  
As You Like It ◽  
A Midsummer Night's Dream ◽  
History Of ◽  
The Way

That Shakespeare adds a limp to the received characterization of Richard III is only the most conspicuous instance of his interest in how actors walked, ran, danced, and wandered. His attention to actors’ footwork, as an originating condition of performance, can be traced from Richard III through A Midsummer Night’s Dream and As You Like It into Macbeth, which is preoccupied with the topic and activity all the way to the protagonist’s melancholy conclusion that ‘Life’s but a walking shadow, a poor player | That struts and frets his hour upon the stage’. Drawing on classical and early modern accounts of how people walk and should walk, on ideas about time and prosody, and the experience of disability, this chapter cites episodes in the history of performance to show how actors, including Alleyn, Garrick, and Olivier, have worked with the opportunities to dramatize footwork that are provided by Shakespeare’s plays.

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