scholarly journals Expression of the Campylobacter jejuni FliD protein and its reaction to chicken sera

2020 ◽  
Vol 367 (14) ◽  
Author(s):  
Xiao-Yan Zhang ◽  
Qian Zhou ◽  
Meng-Jun Tang ◽  
Jun-Hua Pu ◽  
Yan-Feng Fan ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Human Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Capping Protein ◽  
Cultural Status ◽  
Bacterial Gastroenteritis ◽  
Antibody Levels ◽  
Culture Negative

ABSTRACT Campylobacter is a leading causative pathogen of acute bacterial gastroenteritis among humans. Contaminated chicken products are regarded as major sources of human infection. The flagellar capping protein (FliD), which plays important roles in colonization and adhesion to the mucosal surface of chicken ceca, is conserved among Campylobacter jejuni strains. In this study, the recombinant C. jejuni FliD protein was expressed, purified and used as a coated protein to examine the prevalence of C. jejuni antibodies in chickens. The anti-FliD antibody was prevalent among chicken serum samples taken from different farms in the diverse regions of Jiangsu province by using enzyme-linked immunosorbent assay. The Campylobacter antibody was present in culture-negative chickens. No strong dose–response relationships were observed between serum FliD antibody levels and Campylobacter cultural status. These results provide a basis for further evaluating FliD as a vaccine candidate for broiler chickens or for examining host–C. jejuni interactions, with implications for improving food safety.

scholarly journals Characterization and antigenicity of recombinant Campylobacter jejuni flagellar capping protein FliD

2014 ◽  
Vol 63 (4) ◽  
pp. 602-609 ◽  
Author(s):  
Hung-Yueh Yeh ◽  
Kelli L. Hiett ◽  
John E. Line ◽  
Bruce S. Seal
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Nucleotide Sequencing ◽  
Calculated Molecular Mass ◽  
Ionization Time ◽  
Bacterial Flagella ◽  
Capping Protein ◽  
Flight Mass Spectrometry ◽  
Sds Page ◽  
Bacterial Gastroenteritis

Campylobacter jejuni, a flagellated, spiral-rod, Gram-negative bacterium, is the leading pathogen of human acute bacterial gastroenteritis worldwide, and chickens are regarded as a major reservoir of this micro-organism. Bacterial flagella, composed of more than 35 proteins, play important roles in colonization and adhesion to the mucosal surface of chicken caeca. In this study, the flagellar capping protein, FliD, encoded by the fliD gene, from the Campylobacter jenuni D1-39 isolate was expressed and characterized, and its antigenicity determined. The fliD gene comprised 1929 nt, potentially encoding a 642 aa peptide with a calculated molecular mass of 69.6 kDa. This gene was PCR amplified and overexpressed in Escherichia coli. The recombinant FliD protein was purified by cobalt-chelating affinity chromatography and confirmed by nucleotide sequencing of the expression plasmid, SDS-PAGE analysis, His tag detection and matrix-assisted laser desorption/ionization time of flight mass spectrometry. The immunoblot data showed that the purified recombinant FliD protein reacted strongly to sera from broiler chickens older than 4 weeks, indicating that anti-FliD antibody may be prevalent in the poultry population. These results provide a rationale for further evaluation of the FliD protein as a vaccine candidate for broiler chickens to improve food safety for poultry.

scholarly journals Serological Evaluation of Specific-Antibody Levels in Patients Treated for Chronic Chagas' Disease

2007 ◽  
Vol 15 (2) ◽  
pp. 297-302 ◽  
Author(s):  
Olga Sánchez Negrette ◽  
Fernando J. Sánchez Valdéz ◽  
Carlos D. Lacunza ◽  
María Fernanda García Bustos ◽  
María Celia Mora ◽  
...  
Keyword(s):  
Chagas Disease ◽  
Treatment Effectiveness ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Recombinant Antigens ◽  
Antibody Titers ◽  
Laboratory Procedures ◽  
The Individual ◽  
Antibody Levels

ABSTRACT Serological tests are the main laboratory procedures used for diagnosis during the indeterminate and chronic stages of Chagas' disease. A serological regression to negativity is the main criterion used to define parasitological cure in treated patients. The aim of this work was to monitor the individual specificities of antibody levels for 3 years posttreatment in 18 adult patients. Conventional serological techniques (hemagglutination assays and enzyme-linked immunosorbent assay [ELISA]) were modified by using recombinant antigens to detect early markers of treatment effectiveness. For this purpose, serum samples were taken before and during treatment and every 6 months after treatment for at least 3 years. When hemagglutination assays were used, a decrease in antibody levels was observed in only one patient. When ELISA with serum dilutions was used, antibody clearance became much more apparent: in 77.7% (14/18) of the patients, antibody titers became negative with time. This was observed at serum dilutions of 1/320 and occurred between the 6th and the 30th months posttreatment. The immune response and the interval for a serological regression to negativity were different for each patient. For some of the recombinant antigens, only 50% (9/18) of the patients reached the serological regression to negativity. Recombinant antigen 13 might be a good marker of treatment effectiveness, since 66.6% (six of nine) of the patients presented with an early regression to negativity for specific antibodies to this antigen (P = 0.002).

scholarly journals IgM and IgA Antibody Responses in 12 Cases of Human Acquired Toxoplasmosis

1997 ◽  
Vol 39 (6) ◽  
pp. 327-332 ◽  
Author(s):  
Emília E. H. TAKAHASHI ◽  
Cláudio L. ROSSI
Keyword(s):  
Serological Test ◽  
Clinical Manifestations ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igm Antibodies ◽  
Iga Antibodies ◽  
Direct Elisa ◽  
Acute Toxoplasmosis ◽  
Antibody Levels ◽  
Acquired Toxoplasmosis

The persistence, in some subjects, of specific IgM antibodies to Toxoplasma gondii for several months after the acute phase of infection has complicated the interpretation of serological test results for toxoplasmosis. Several reports have emphasized the value of the detection of Toxoplasma-specific IgA antibodies for the diagnosis of acute toxoplasmosis. In this article, we report the follow-up profiles of Toxoplasma-specific IgM and IgA antibodies in serum samples obtained from 12 patients at various intervals after the onset of the clinical manifestations of infection. IgM antibodies were detected by the indirect immunofluorescence (IIF) test, antibody capture enzyme-linked immunosorbent assay (cELISA) and enzyme-mediated chemilluminescent technique (CmL). IgA antibodies were quantified by the direct ELISA (dELISA) and cELISA procedures. As defined by the manufacturer of the cELISA test for IgA used, most patients with acute toxoplasmosis have antibody levels > 40 arbritary units per ml (AU/ml). At values > 40 AU/ml, the cELISA for IgA detected significant antibody levels for a shorter time than the other techniques used for IgM and IgA detection. However, IgA levels <FONT FACE="Symbol">£</font> 40 AU/ml do not exclude the possibility of acute toxoplasmosis since such levels can be reached very soon after infection with T. gondii. The results obtained in the present study show that the serological diagnosis of acute toxoplasmosis may not be such an easy task. Our data suggest that use of the IgA-cELISA concomitantly with IgM antibody screening could permit, in some circumstances, a more efficient diagnosis of acute acquired toxoplasmosis

scholarly journals Seroprevalence of avian metapneumovirus infection in broiler and broiler breeder chickens in Iran

2011 ◽  
Vol 56 (No. 8) ◽  
pp. 395-399 ◽  
Author(s):  
M. Rahimi
Keyword(s):  
Broiler Chickens ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Upper Respiratory Tract ◽  
Serum Samples ◽  
Broiler Breeder ◽  
Blood Samples ◽  
Avian Metapneumovirus ◽  
Commercial Chicken ◽  
Future Work

Avian metapneumovirus causes an acute highly contagious upper respiratory tract infection primarily of turkeys and chickens. The disease can cause significant economic losses in turkey and chicken flocks, particularly when exacerbated by secondary pathogens. The purpose of this study was to determine the prevalence of avian metapneumovirus antibodies in broiler and broiler breeder flocks in Kermanshah province, west of Iran. All the flocks had not been vaccinated against avian metapneumovirus. The province were divided into four geographic areas; southwest, southeast, northwest, and northeast. Flocks in each area, and 14&ndash;15 birds in each flock, were randomly sampled. The blood samples were taken regardless of the presence of any signs of respiratory or any other clinical disease in the flocks. A total of 435 blood samples were collected from 30 commercial chicken flocks (24 broiler flocks, aged between six and eight weeks, and six broiler breeder flocks, aged between 56 and 72 weeks). The presence of antibodies against avian metapneumovirus in each serum sample was tested twice by enzyme-linked immunosorbent assay using a commercial kit which was able to determine antibodies against A, B and C subtypes of avian metapneumovirus. Out of 347 serum samples obtained from broiler chickens, 167 (48.1%) were positive to avian metapneumovirus antibodies, which represented 20 (83.3%) of 24 examined broiler flocks. Out of 88 samples obtained from broiler breeder chickens, 82 (93.2%) were positive to avian metapneumovirus antibodies, which belonged to six (100%) of examined broiler breeder flocks. Detection of anti-avian metapneumovirus antibodies among broiler breeder (100%) was higher than broiler (83.3%) flocks. A higher rate of seropositivity (83.3% of samples and 100% of broiler flocks) was observed in northwest. The results of this study may indicate the possible involvement of avian metapneumovirus in the respiratory disease we are seeing in chickens in Iran. Its prevalence has to be investigated in other parts of Iran. Future work may and should include the use of molecular methods and isolation of the virus. Isolation of avian metapneumovirus will allow the possibility of making autogenous vaccines.

scholarly journals Tetanus Immunity among Women Aged 15 to 39 Years in Cambodia: a National Population-Based Serosurvey, 2012

2016 ◽  
Vol 23 (7) ◽  
pp. 546-554 ◽  
Author(s):  
Heather M. Scobie ◽  
Bunsoth Mao ◽  
Sokhal Buth ◽  
Kathleen A. Wannemuehler ◽  
Charlotte Sørensen ◽  
...  
Keyword(s):  
High Sensitivity ◽  
Population Based ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cluster Sampling ◽  
Serum Samples ◽  
Nulliparous Women ◽  
Vaccination Campaigns ◽  
Antibody Levels ◽  
Multiple Pathogens ◽  
Tetanus Immunity

To monitor progress toward maternal and neonatal tetanus elimination (MNTE) in Cambodia, we conducted a nationwide serosurvey of tetanus immunity in 2012. Multistage cluster sampling was used to select 2,154 women aged 15 to 39 years. Tetanus toxoid antibodies in serum samples were measured by gold-standard double-antigen enzyme-linked immunosorbent assay (DAE) and a novel multiplex bead assay (MBA). Antibody concentrations of ≥0.01 IU/ml by DAE or the equivalent for MBA were considered seroprotective. Estimated tetanus seroprotection was 88% (95% confidence interval [CI], 86 to 89%); 64% (95% CI, 61 to 67%) of women had antibody levels of ≥1.0 IU/ml. Seroprotection was significantly lower (P< 0.001) among women aged 15 to 19 years (63%) and 20 to 24 years (87%) than among those aged ≥25 years (96%), among nulliparous women than among parous women (71 versus 97%), and among those living in the western region than among those living in other regions (82 versus 89%). The MBA showed high sensitivity (99% [95% CI, 98 to 99%]) and specificity (92% [95% CI, 88 to 95%]) compared with DAE. Findings were compatible with MNTE in Cambodia (≥80% protection). Tetanus immunity gaps should be addressed through strengthened routine immunization and targeted vaccination campaigns. Incorporating tetanus testing in national serosurveys using MBAs, which can measure immunity to multiple pathogens simultaneously, may be beneficial for monitoring MNTE.

scholarly journals Bacteriophage Therapy To Reduce Campylobacter jejuni Colonization of Broiler Chickens

2005 ◽  
Vol 71 (11) ◽  
pp. 6554-6563 ◽  
Author(s):  
C. Loc Carrillo ◽  
R. J. Atterbury ◽  
A. El-Shibiny ◽  
P. L. Connerton ◽  
E. Dillon ◽  
...  
Keyword(s):  
Campylobacter Jejuni ◽  
Broiler Chickens ◽  
Phage Therapy ◽  
Experimental Models ◽  
Human Infection ◽  
Lytic Bacteriophage ◽  
Bacteriophage Therapy ◽  
Bacteriophage Infection ◽  
Low Passage

ABSTRACT Colonization of broiler chickens by the enteric pathogen Campylobacter jejuni is widespread and difficult to prevent. Bacteriophage therapy is one possible means by which this colonization could be controlled, thus limiting the entry of campylobacters into the human food chain. Prior to evaluating the efficacy of phage therapy, experimental models of Campylobacter colonization of broiler chickens were established by using low-passage C. jejuni isolates HPC5 and GIIC8 from United Kingdom broiler flocks. The screening of 53 lytic bacteriophage isolates against a panel of 50 Campylobacter isolates from broiler chickens and 80 strains isolated after human infection identified two phage candidates with broad host lysis. These phages, CP8 and CP34, were orally administered in antacid suspension, at different dosages, to 25-day-old broiler chickens experimentally colonized with the C. jejuni broiler isolates. Phage treatment of C. jejuni-colonized birds resulted in Campylobacter counts falling between 0.5 and 5 log10 CFU/g of cecal contents compared to untreated controls over a 5-day period postadministration. These reductions were dependent on the phage-Campylobacter combination, the dose of phage applied, and the time elapsed after administration. Campylobacters resistant to bacteriophage infection were recovered from phage-treated chickens at a frequency of <4%. These resistant types were compromised in their ability to colonize experimental chickens and rapidly reverted to a phage-sensitive phenotype in vivo. The selection of appropriate phage and their dose optimization are key elements for the success of phage therapy to reduce campylobacters in broiler chickens.

The effect of plasma exchange on serum anti-JC virus antibodies

2012 ◽  
Vol 19 (7) ◽  
pp. 912-919 ◽  
Author(s):  
Meena Subramanyam ◽  
Tatiana Plavina ◽  
Bhupendra O Khatri ◽  
Robert J Fox ◽  
Susan E Goelz
Keyword(s):  
Plasma Exchange ◽  
Immune Reconstitution ◽  
Jc Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Fold Reduction ◽  
The Central Nervous System ◽  
Igg Isotypes ◽  
Circulating Antibodies ◽  
Antibody Levels

Objective: Natalizumab, a highly effective treatment for multiple sclerosis (MS) and Crohn’s disease, is associated with progressive multifocal leukoencephalopathy (PML). Upon suspicion or diagnosis of PML, plasma exchange (PLEX) is performed to remove natalizumab from the circulation, allowing immune reconstitution of the central nervous system. Since PLEX may also remove other circulating antibodies, we examined the effects of PLEX on serum immunoglobulin (IgG) and anti–JC virus (JCV) antibody levels in MS patients with and without PML. Methods: Serum samples from 12 natalizumab-treated patients without PML collected before, during and after PLEX were tested for IgG isotypes using a commercial assay, and for anti-JCV antibodies using a two-step enzyme-linked immunosorbent assay. Five natalizumab-treated PML patients who underwent PLEX were also tested for anti-JCV antibodies. Results: PLEX produced a two- to three-fold reduction in all IgG isotypes. Among patients without PML, 42% (five of 12 patients) had detectable anti-JCV antibodies before PLEX; in these patients, anti-JCV antibodies were reduced approximately two- to five-fold, with levels returning to 50–100 percent of baseline two weeks after the final PLEX. The five PML patients, all of whom had detectable anti-JCV antibodies before PLEX, experienced similar reductions in anti-JCV antibody levels following PLEX. Conclusions: Our results indicate that PLEX effectively removes circulating antibodies; however, levels of endogenous anti-JCV antibody, unlike exogenously administered natalizumab, were replenished relatively quickly following PLEX. While interpretation of anti-JCV antibody levels during or within two weeks after PLEX may be problematic, humoral JCV immunity is not abolished by PLEX and antibody levels are rapidly restored.

scholarly journals Serological survey of Coxiella burnetii infections in dairy cattle, sheep, goats and zoo animals in Hungary – Short communication

2021 ◽  
Author(s):  
Attila Dobos ◽  
István Fodor ◽  
Gerda Kiss ◽  
Miklós Gyuranecz
Keyword(s):  
Dairy Cattle ◽  
Coxiella Burnetii ◽  
Large Scale ◽  
Q Fever ◽  
Small Ruminants ◽  
Human Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Ruminant Species ◽  
Zoo Animals

AbstractQ fever is a disease of high zoonotic potential, but interest in its causative agent is rather low although it causes some public health problems in Hungary. The prevalence of Q fever is highly variable by country. The main reservoirs of the disease are the same domestic ruminant species everywhere, but the epidemiological profile depends on the features of the specific reservoir. The aim of this large-scale study was to demonstrate the importance of Q fever in different species as a possible source for human infection in most regions of Hungary. A total of 851 serum samples from 44 dairy farms, 16 sheep flocks, 4 goat farms and 3 zoos located in different parts of Hungary were tested. The presence of antibodies to Coxiella burnetii was surveyed in dairy cattle (n = 547), goats (n = 71), sheep (n = 200) and zoo animals (n = 33). The animal species tested in Hungary showed different seroprevalence values of C. burnetii infection. Seropositivity by the enzyme-linked immunosorbent assay was found in 258 out of 547 (47.2%) cows and in 69 out of 271 (25.5%) small ruminants, among them in 47 out of 200 (23.5%) sheep and in 22 out of 71 (31.0%) goats. Antibodies to C. burnetii were not detected in zoo animals. Seropositivity was demonstrated in 44 out of 44 (100%) dairy cattle farms, with at least one serum sample found to be positive on each farm. The seropositivity rate of small ruminant farms was 55.0% (11 positive out of 20 tested), with 9 out of 16 (56.3%) sheep flocks and 2 out of 4 (50.0%) goat herds showing seropositivity.

scholarly journals Immunoglobulin G and A Antibody Responses to Bacteroides forsythus and Prevotella intermedia in Sera and Synovial Fluids of Arthritis Patients

2003 ◽  
Vol 10 (6) ◽  
pp. 1043-1050 ◽  
Author(s):  
Ketil Moen ◽  
Johan G. Brun ◽  
Tor Magne Madland ◽  
Turid Tynning ◽  
Roland Jonsson
Keyword(s):  
Immune Responses ◽  
Immunoglobulin G ◽  
Enzyme Linked Immunosorbent Assay ◽  
Prevotella Intermedia ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Igg Antibody ◽  
Iga Antibodies ◽  
Iga Antibody ◽  
Antibody Levels

ABSTRACT The objective of the present study was to investigate immunoglobulin G (IgG) and IgA antibody immune responses to Porphyromonas gingivalis, Prevotella intermedia, Bacteroides forsythus, and Candida albicans in the sera of patients with rheumatoid arthritis (RA), the synovial fluid (SF) of patients with RA (RA-SF samples), and the SF of patients without RA (non-RA-SF samples). An enzyme-linked immunosorbent assay was used to determine IgG and IgA antibody levels in 116 serum samples from patients with RA, 52 RA-SF samples, and 43 non-RA-SF samples; and these were compared with those in SF samples from 9 patients with osteoarthritis (OA-SF samples) and the blood from 100 donors (the control [CTR] group). Higher levels of IgG antibodies against B. forsythus (P < 0.0001) and P. intermedia (P < 0.0001) were found in non-RA-SF samples than in OA-SF samples, and higher levels of IgG antibodies against B. forsythus (P = 0.003) and P. intermedia (P = 0.024) were found in RA-SF samples than in OA-SF samples. Significantly higher levels of IgA antibodies against B. forsythus were demonstrated in both RA-SF and non-RA-SF samples than in OA-SF samples. When corrected for total Ig levels, levels of IgG antibody against B. forsythus were elevated in RA-SF and non-RA-SF samples compared to those in OA-SF samples. Lower levels of Ig antibodies against B. forsythus were found in the sera of patients with RA than in the plasma of the CTR group for both IgG (P = 0.003) and IgA (P < 0.0001). When corrected for total Ig levels, the levels of IgG and IgA antibodies against B. forsythus were still found to be lower in the sera from patients with RA than in the plasma of the CTR group (P < 0.0001). The levels of antibodies against P. gingivalis and C. albicans in the sera and SF of RA and non-RA patients were comparable to those found in the respective controls. The levels of IgG and IgA antibodies against B. forsythus were elevated in SF from patients with RA and non-RA-SF samples compared to those in OA-SF samples. Significantly lower levels of IgG and IgA antibodies against B. forsythus were found in the sera of patients with RA than in the plasma of the CTR group. This indicates the presence of an active antibody response in synovial tissue and illustrates a potential connection between periodontal and joint diseases.

Dot immunogold filtration assay (DIGFA) for the rapid detection of specific antibodies against the rat lungwormAngiostrongylus cantonensis(Nematoda: Metastrongyloidea) using purified 31-kDa antigen

2013 ◽  
Vol 88 (4) ◽  
pp. 396-401 ◽  
Author(s):  
P. Eamsobhana ◽  
X.X. Gan ◽  
A. Ma ◽  
Y. Wang ◽  
D. Wanachiwanawin ◽  
...  
Keyword(s):  
Large Scale ◽  
Protein A ◽  
Epidemiological Studies ◽  
Human Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Parasitic Diseases ◽  
Serum Samples ◽  
Specific Antibodies ◽  
North East ◽  
Gold Conjugate

AbstractA rapid dot immunogold filtration assay (DIGFA) was adopted for specific immunodiagnosis of human cerebral angiostrongyliasis, using purified 31-kDa glycoprotein specific toAngiostrongylus cantonensisas diagnostic antigen and protein A colloidal gold conjugate as antigen–antibody detector. A total of 59 serum samples were assayed – 11 samples from clinically diagnosed patients with detectableA. cantonensis-specific antibody in immunoblotting; 23 samples from patients with other related parasitic diseases, i.e. gnathostomiasis (n= 8), cysticercosis (n= 5), toxocariasis (n= 2), filariasis (n= 4), paragonimiasis (n= 2) and malaria (n= 2); and 25 samples from normal healthy subjects. The sensitivity and specificity of DIGFA to detect anti-A. cantonensisspecific antibodies in serologically confirmed angiostrongyliasis cases, were both 100%. No positive DIGFA was observed in cases with other parasitic diseases, and the healthy control subjects. The 3-min DIGFA is as sensitive and specific as the 3-h immunoblot test in angiostrongyliasis confirmed cases that revealed a 31-kDa reactive band. The gold-based DIGFA is more rapid and easier to perform than the traditional enzyme-linked immunosorbent assay (ELISA). The test utilizing purifiedA.cantonensisantigen is reliable and reproducible for specific immunodiagnosis of human infection withA. cantonensis– thus can be applied as an additional routine test for clinical diagnostic support. Large-scale sero-epidemiological studies in endemic communities in north-east Thailand are under way to evaluate its usefulness under field conditions.

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