scholarly journals Use of recombinant Brucella outer membrane proteins 19, 25, and 31 for serodiagnosis of bovine brucellosis

2020 ◽  
Vol 13 (7) ◽  
pp. 1439-1447
Author(s):  
Aitbay Bulashev ◽  
Orken Akibekov ◽  
Alfiya Syzdykova ◽  
Zhanbolat Suranshiyev ◽  
Bakytkali Ingirbay
Keyword(s):  
Outer Membrane ◽  
Single Injection ◽  
Outer Membrane Proteins ◽  
Enzyme Linked Immunosorbent Assay ◽  
Post Inoculation ◽  
Serological Testing ◽  
Bovine Brucellosis ◽  
Serum Samples ◽  
Serological Tests ◽  
Time Point

Background and Aim: Brucellosis remains one of the most common zoonoses. The current anti-brucellosis measures are largely deemed ineffective due to a lack of specificity of conventional serological tests. This study evaluated the use of Brucella outer membrane protein (Omp)19 for serodiagnostic testing. Materials and Methods: The antigenicity of recombinant Brucella Omp19, Omp25, and Omp31 was examined in serum samples from mice and rabbits immunized with Omp19 or Brucella abortus 19 whole cell (WC) and 12 and 152 cows experimentally or naturally infected with brucellosis, respectively. Serum samples were collected from 151 cows that were vaccinated with B. abortus 19 and 12 unvaccinated heifers that were maintained on a brucellosis-free farm. Results: Immunization with Omp19 resulted in antibody production in mice after a single injection without the use of adjuvant. Serum antibodies obtained from rabbits immunized with inactivated B. abortus strain 19 WC targeted Omps by enzyme-linked immunosorbent assay (ELISA) and Western blot. Antibodies targeting Omp19 were identified in all B. abortus strain 544 experimentally infected cows at day 14 post-inoculation (p.i.); Omp25 was detected by ELISA at day 28 p.i., while an ELISA targeting Omp31 was negative for 25% of cows at this time point. Omp19 and Omp25 were readily detected by sera from cows from a new epizootic focus. Antibodies recognizing Omps were also detected in >50% of the animals maintained in a brucellosis-free herd at 10 months after vaccination. Conclusion: Brucella Omp19 in combination with Omp25 and Omp31 may be utilized as target antigens in an ELISA designed for serological testing of unvaccinated cattle.

scholarly journals Evaluation of chimeric proteins for serological diagnosis of brucellosis in cattle

2021 ◽  
pp. 2187-2196
Author(s):  
Aitbay K. Bulashev ◽  
Bakytkali K. Ingirbay ◽  
Kanatbek N. Mukantayev ◽  
Alfiya S. Syzdykova
Keyword(s):  
Membrane Proteins ◽  
Outer Membrane ◽  
Outer Membrane Proteins ◽  
Cross Reactivity ◽  
Accurate Diagnosis ◽  
Expression Vectors ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chimeric Proteins ◽  
Serum Samples ◽  
Serological Tests

Background and Aim: An accurate diagnosis of Brucella-infected animals is one of the critical measures in eradication programs. Conventional serological tests based on whole-cell (WC) antigens and detecting antibodies against pathogen-associated lipopolysaccharide might give false-positive results due to the cross-reactivity with other closely related bacteria. This study evaluated the serological potential of Brucella spp. chimeric outer membrane proteins (Omps) as antigens in an indirect enzyme-linked immunosorbent assay (i-ELISA). Materials and Methods: The chimeric gene constructs of the most immunodominant regions of Brucella Omps 25+31, 25+19, and 19+31 were cloned into the pET28a expression vectors and transformed into Escherichia coli BL21 (DE3). The serological potential of chimeric proteins compared with single recombinant Omps (rOmps)19, 25, and/or 31 were studied on blood serum samples of (i) a rabbit immunized with killed Brucella abortus 19WC, (ii) mice immunized with single rOmps, (iii) cows seropositive for brucellosis by rose Bengal test, and (iv) cattle naturally and/or experimentally infected with brucellosis. Results: E. coli BL21 actively produced Brucella chimeric rOmps, the concentration of which reached a maximum level at 6 h after isopropyl-β-D-1-thiogalactopyranoside stimulation. Target proteins were antigenic and expressed in an active state, as recognized by rabbit anti-B. abortus antibodies in an i-ELISA and western blotting. Murine antibodies against the single rOmps reacted with chimeric antigens, and conversely, antichimeric antibodies found their epitopes in single proteins. Brucella chimeric rOmps showed higher antigenicity in blood sera of seropositive cattle kept in the hotbed of the infection and/or experimentally challenged with brucellosis than single proteins. Conclusion: Brucella chimeric recombinant outer membrane proteins could be a potential antigen candidate for developing an ELISA test for accurate diagnosis of bovine brucellosis.

Comparison of indirect haemagglutination test, gel-diffusion precipitin test, and enzyme-linked immunosorbent assay for detection of serum antibodies to Pasteurella multocida in naturally and experimentally infected rabbits

1994 ◽  
Vol 28 (1) ◽  
pp. 19-25 ◽  
Author(s):  
Eiichi Kawamoto ◽  
Takuo Sawada ◽  
Toru Sato ◽  
Kiyoshi Suzuki ◽  
Tsutomu Maruyama
Keyword(s):  
Immunosorbent Assay ◽  
Pasteurella Multocida ◽  
Rabbit Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Post Inoculation ◽  
Serum Samples ◽  
Serological Tests ◽  
Indirect Haemagglutination ◽  
Precipitin Test ◽  
Gel Diffusion

Enzyme-linked immunosorbent assay (ELISA), gel-diffusion precipitin test (GDPT), and indirect haem agglutination test (IHAT) were evaluated for the detection of antibodies to Pasteurella multocida in both naturally and experimentally infected rabbits. A total of 285 rabbit serum samples from 7 rabbit colonies were tested by ELISA, GDPT, and IHAT, and nasal cultures were taken coincidentally to use as the standard in the serological tests. There was better correlation (98.0%) between the results of ELISA and positive nasal culture than between the GDPT (86.3%) or IHAT (23.5%) and positive nasal culture. In addition, ELISA and GDPT were positive in 26 (11.1%) and 21 (9.0%) of 234 serum samples from nasal culture negative rabbits, respectively. In experimentally infected rabbits, antibodies detected by the ELISA and GDPT began to rise one to 3 weeks post-inoculation. IHAT did not detect antibodies. These results are discussed in terms of value to serodiagnosis of rabbit pasteurellosis.

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 Serodiagnosis of Louse-Borne Relapsing Fever with Glycerophosphodiester Phosphodiesterase (GlpQ) fromBorrelia recurrentis

2000 ◽  
Vol 38 (10) ◽  
pp. 3561-3571 ◽  
Author(s):  
Stephen F. Porcella ◽  
Sandra J. Raffel ◽  
Merry E. Schrumpf ◽  
Martin E. Schriefer ◽  
David T. Dennis ◽  
...  
Keyword(s):  
Serological Test ◽  
Geometric Mean ◽  
Eastern Africa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Relapsing Fever ◽  
Serum Samples ◽  
Serological Tests ◽  
Whole Cell ◽  
Convalescent Phase ◽  
Glycerophosphodiester Phosphodiesterase

Human louse-borne relapsing fever occurs in sporadic outbreaks in central and eastern Africa that are characterized by significant morbidity and mortality. Isolates of the causative agent,Borrelia recurrentis, were obtained from the blood of four patients during a recent epidemic of the disease in southern Sudan. TheglpQ gene, encoding glycerophosphodiester phosphodiesterase, from these isolates was sequenced and compared with the glpQ sequences obtained from other relapsing-fever spirochetes. Previously we showed that GlpQ of Borrelia hermsii is an immunogenic protein with utility as a serological test antigen for discriminating tick-borne relapsing fever from Lyme disease. In the present work, we cloned and expressed theglpQ gene from B. recurrentis and used recombinant GlpQ in serological tests. Acute- and convalescent-phase serum samples obtained from 42 patients with louse-borne relapsing fever were tested with an indirect immunofluorescence assay (IFA) and an enzyme-linked immunosorbent assay (ELISA) that used whole cells ofB. recurrentis and with immunoblotting to whole-cell lysates of the spirochete and Escherichia coli producing recombinant GlpQ. The geometric mean titers of the acute- and convalescent-phase serum samples measured by IFA were 1:83 and 1:575, respectively. The immunoblot analysis identified a high level of reactivity and seroconversion to GlpQ, and the assay was more sensitive than the whole-cell IFA and ELISA using purified, recombinant histidine-tagged GlpQ. Serum antibodies to GlpQ and other antigens persisted for 27 years in one patient. We conclude that assessment of anti-GlpQ antibodies will allow serological confirmation of louse-borne relapsing fever and determination of disease prevalence.

scholarly journals Bayesian evaluation of three serological tests for the diagnosis of bovine brucellosis in Bangladesh

2019 ◽  
Vol 147 ◽  
Author(s):  
A. K. M. A. Rahman ◽  
S. Smit ◽  
B. Devleesschauwer ◽  
P. Kostoulas ◽  
E. Abatih ◽  
...  
Keyword(s):  
Latent Class ◽  
Latent Class Model ◽  
Dairy Farm ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Brucellosis ◽  
Class Model ◽  
Serological Tests ◽  
Predictive Values ◽  
Conditional Dependence

AbstractWe evaluated the performance of three serological tests – an immunoglobulin G indirect enzyme linked immunosorbent assay (iELISA), a Rose Bengal test and a slow agglutination test (SAT) – for the diagnosis of bovine brucellosis in Bangladesh. Cattle sera (n = 1360) sourced from Mymensingh district (MD) and a Government owned dairy farm (GF) were tested in parallel. We used a Bayesian latent class model that adjusted for the conditional dependence among the three tests and assumed constant diagnostic accuracy of the three tests in both populations. The sensitivity and specificity of the three tests varied from 84.6% to 93.7%, respectively. The true prevalences of bovine brucellosis in MD and the GF were 0.6% and 20.4%, respectively. Parallel interpretation of iELISA and SAT yielded the highest negative predictive values: 99.9% in MD and 99.6% in the GF; whereas serial interpretation of both iELISA and SAT produced the highest positive predictive value (PPV): 99.9% in the GF and also high PPV (98.9%) in MD. We recommend the use of both iELISA and SAT together and serial interpretation for culling and parallel interpretation for import decisions. Removal of brucellosis positive cattle will contribute to the control of brucellosis as a public health risk in Bangladesh.

scholarly journals Evaluation of the Serologic Cross-Reactivity between Transmissible Gastroenteritis Coronavirus and Porcine Respiratory Coronavirus Using Commercial Blocking Enzyme-Linked Immunosorbent Assay Kits

mSphere ◽  
2019 ◽  
Vol 4 (2) ◽  
Author(s):  
Ronaldo Magtoto ◽  
Korakrit Poonsuk ◽  
David Baum ◽  
Jianqiang Zhang ◽  
Qi Chen ◽  
...  
Keyword(s):  
Culture Medium ◽  
Large Deletion ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Transmissible Gastroenteritis Virus ◽  
Serum Samples ◽  
Serological Tests ◽  
Transmissible Gastroenteritis ◽  
Respiratory Coronavirus ◽  
Porcine Respiratory

ABSTRACTThis study compared the performances of three commercial transmissible gastroenteritis virus/porcine respiratory coronavirus (TGEV/PRCV) blocking enzyme-linked immunosorbent assays (ELISAs) using serum samples (n = 528) collected over a 49-day observation period from pigs inoculated with TGEV strain Purdue (n = 12), TGEV strain Miller (n = 12), PRCV (n= 12), or with virus-free culture medium (n = 12). ELISA results were evaluated both with “suspect” results interpreted as positive and then as negative. All commercial kits showed excellent diagnostic specificity (99 to 100%) when testing samples from pigs inoculated with virus-free culture medium. However, analyses revealed differences between the kits in diagnostic sensitivity (percent TGEV- or PRCV-seropositive pigs), and all kits showed significant (P < 0.05) cross-reactivity between TGEV and PRCV serum antibodies, particularly during early stages of the infections. Serologic cross-reactivity between TGEV and PRCV seemed to be TGEV strain dependent, with a higher percentage of PRCV-false-positive results for pigs inoculated with TGEV Purdue than for TGEV Miller. Moreover, the overall proportion of false positives was higher when suspect results were interpreted as positive, regardless of the ELISA kit evaluated.IMPORTANCECurrent measures to prevent TGEV from entering a naive herd include quarantine and testing for TGEV-seronegative animals. However, TGEV serology is complicated due to the cross-reactivity with PRCV, which circulates subclinically in most swine herds worldwide. Conventional serological tests cannot distinguish between TGEV and PRCV antibodies; however, blocking ELISAs using antigen containing a large deletion in the amino terminus of the PRCV S protein permit differentiation of PRCV and TGEV antibodies. Several commercial TGEV/PRCV blocking ELISAs are available, but performance comparisons have not been reported in recent research. This study demonstrates that the serologic cross-reactivity between TGEV and PRCV affects the accuracy of commercial blocking ELISAs. Individual test results must be interpreted with caution, particularly in the event of suspect results. Therefore, commercial TGEV/PRCV blocking ELISAs should only be applied on a herd basis.

scholarly journals Human herpes virus 8 antibodies in HIV-positive patients in Surabaya, Indonesia

2020 ◽  
Author(s):  
Devi Oktafiani ◽  
Ni Luh Ayu Megasari ◽  
Elsa Fitriana ◽  
Nasronudin ◽  
Maria Inge Lusida ◽  
...  
Keyword(s):  
Drug Users ◽  
Sandwich Elisa ◽  
Human Herpesvirus ◽  
Intravenous Drug ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hiv Positive ◽  
Intravenous Drug Users ◽  
Serum Samples ◽  
Serological Tests ◽  
Human Herpes Virus 8

Background: Human herpesvirus 8 (HHV-8) infection is etiologically related to Kaposi’s sarcoma. Antibodies directed against HHV-8 can be detected in 80%–95% of HIV-seropositive patients with KS. HHV-8 serological tests have been done in several countries in Southeast Asia such as Malaysia, and Thailand however no serological data is available in Indonesia. This study was to examine the presence of HHV-8 antibodies in HIV-positive patients in Surabaya, Indonesia. Material and methods: Ninety-one serum samples were collected from HIV-positive patients in Surabaya, Indonesia. Human immunodeficiency virus-positive serum samples were collected from 10 homosexual men, 25 intravenous drug users (IVDUs) and 56 heterosexuals. Serums were then tested for the presence of HHV-8 antibody by using sandwich ELISA (Abbexa Ltd, Cambridge, UK). Results: The total of 91 HIV-infected were testing with antibodies to HHV-8 using enzyme-linked immunosorbent assay. Antibodies of HHV-8 were detected in 7/91 (7.7%) of the samples. According to a gender, six men (85.7%) and a women (14.3%) were positive of HHV-8 antibodies. No correlation regarding the gender and age from this study. The antibodies of HHV-8 was detected among intravenous drug users (IVDUs) men 5/7 (42.8%) and 2/7 (28.6%) from homosexual and heterosexual, respectively. Conclusion: This study found the presence of HHV-8 antibodies in 7.7% of patients in Surabaya, Indonesia. This finding was higher more than Southeast Asian countries. The patients with a positive result could suggest measures to prevent HHV-8 infection.

scholarly journals Detection of bluetongue virus antibodies in sheep from Paraná, Brazil

2020 ◽  
Vol 41 (3) ◽  
pp. 879
Author(s):  
Maria Carolina Ricciardi Sbizera ◽  
Luiz Fernando Coelho da Cunha Filho ◽  
Michele Lunardi ◽  
Simone Fernanda Nedel Pertile ◽  
Thais Helena Constantino Patelli ◽  
...  
Keyword(s):  
Bluetongue Virus ◽  
Animal Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
Serum Samples ◽  
Agar Gel ◽  
Serological Tests ◽  
High Occurrence ◽  
Predominant Factor ◽  
World Organization

Bluetongue (BT) is an infectious and non-contagious disease caused by bluetongue virus (BTV) belonging to the genus Orbivirus. It is transmitted by a hematophagous vector, Culicoides sp., to ruminants, particularly to sheep, which are most susceptible to this disease. The main serological tests are agar gel immunodiffusion (AGID), which is recommended by the World Organization for Animal Health (OIE), and the competitive enzyme-linked immunosorbent assay (cELISA), which has the advantage of no cross-reaction with other orbiviruses. The aim was to compare the results of these two tests by conducting them on sera collected from sheep in the state of Paraná, Brazil. From March to October 2017, serum samples were collected from 270 sheep from 10 farms in six mesoregions of Paraná. The samples were subjected to AGID and cELISA to detect antibodies against BTV. Based on the test results, we classified the sheep as low, moderate, and high occurrence. The results demonstrated that 64.81% (175/270) of the sheep were seropositive through the cELISA test, showing a high occurrence, and 41.11% (111/270) were seropositive through the AGID test, indicating a moderate occurrence. The concordance between the tests was moderate (0.51) as determined by the Kappa coefficient. Among the studied farms, 90% (9/10) presented at least one seropositive sheep, and the number of animals tested positive by the cELISA test was higher than those by the AGID test. Favorable climate, which favors the presence and multiplication of the culicoid vector and the occurrence of infection, was the biggest predominant factor responsible for the obtained results. The low occurrence in farms with milder climate suggest that the presence of antibodies also occurs due to the low pathogenicity of circulating serotypes in the different mesoregions studied. It is concluded that BTV infection is present in the sheep herds in Paraná, and the occurrence was moderate detected by AGID test and high detected by cELISA test.

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