scholarly journals Serological Diagnosis of Brucella Infections in Odontocetes

2009 ◽  
Vol 16 (6) ◽  
pp. 906-915 ◽  
Author(s):  
Gabriela Hernández-Mora ◽  
Charles A. Manire ◽  
Rocío González-Barrientos ◽  
Elías Barquero-Calvo ◽  
Caterina Guzmán-Verri ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Brucella Melitensis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dot Blot ◽  
Serum Samples ◽  
Igg Antibody ◽  
Serological Tests ◽  
Diagnostic Potential ◽  
Control Serum ◽  
History Of

ABSTRACT Brucella ceti causes disease in Odontoceti. The absence of control serum collections and the diversity of cetaceans have hampered the standardization of serological tests for the diagnosis of cetacean brucellosis. Without a “gold” standard for sensitivity and specificity determination, an alternative approach was followed. We designed an indirect enzyme-linked immunosorbent assay (iELISA) that recognizes immunoglobulins G (IgGs) from 17 odontocete species as a single group. For the standardization, we used Brucella melitensis and Brucella abortus lipopolysaccharides, serum samples from seven resident odontocetes with no history of infectious disease displaying negative rose bengal test (RBT) reactions, and serum samples from seven dolphins infected with B. ceti. We compared the performance of the iELISA with those of the protein G ELISA (gELISA), the competitive ELISA (cELISA), and the immunofluorescence (IF) and dot blot (DB) tests, using 179 odontocete serum samples and RBT as the reference. The diagnostic potential based on sensitivity and specificity of the iELISA was superior to that of gELISA and cELISA. The correlation and agreement between the iELISA and the gELISA were relatively good (R i/g 2 = 0.65 and κi/g = 0.66, respectively), while the correlation and agreement of these two ELISAs with cELISA were low (R i/c 2 = 0.46, R g/c 2 = 0.37 and κi/c = 0.62, κg/c = 0.42). In spite of using the same anti-odontocete IgG antibody, the iELISA was more specific than were the IF and DB tests. An association between high antibody titers and the presence of neurological symptoms in dolphins was observed. The prediction is that iELISA based on broadly cross-reacting anti-dolphin IgG antibody would be a reliable test for the diagnosis of brucellosis in odontocetes, including families not covered in this study.

scholarly journals Evaluation of the performance of three serological tests for diagnosis of Leishmania infantum infection in dogs using latent class analysis

2020 ◽  
Vol 29 (4) ◽  
Author(s):  
Asier Basurco ◽  
Alda Natale ◽  
Katia Capello ◽  
Antonio Fernández ◽  
María Teresa Verde ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Leishmania Infantum ◽  
Latent Class ◽  
Rapid Test ◽  
Antibody Test ◽  
Latent Class Models ◽  
Canine Leishmaniasis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests

Abstract Canine leishmaniasis (CanL) is a disease caused by Leishmania infantum. Serological methods are the most common diagnostic techniques used for the diagnosis of the CanL. The objective of our study was to estimate the sensitivity and specificity of one in-house ELISA kit (ELISA UNIZAR) and three commercially available serological tests (MEGACOR Diagnostik GmbH) including an immunochromatographic rapid test (FASTest LEISH®), an immunofluorescent antibody test (MegaFLUO LEISH®) and an enzyme-linked immunosorbent assay (MegaELISA LEISH®), using latent class models in a Bayesian analysis. Two hundred fifteen serum samples were included. The highest sensitivity was achieved for FASTest LEISH® (99.38%), ELISA UNIZAR (99.37%), MegaFLUO LEISH® (99.36%) followed by MegaELISA LEISH® (98.49%). The best specificity was obtained by FASTest LEISH® (98.43%), followed by ELISA UNIZAR (97.50%), whilst MegaFLUO LEISH® and MegaELISA LEISH® obtained the lower specificity (91.94% and 91.93%, respectively). The results of present study indicate that the immunochromatographic rapid test evaluated FASTest LEISH® show similar levels of sensitivity and specificity to the quantitative commercial tests. Among quantitative serological tests, sensitivity and specificity were similar considering ELISA or IFAT techniques.

scholarly journals Comparative Analysis of the Serological Reactivity of Individuals with Clinical History of Malaria using Two Different ELISA Tests

Diagnostics ◽  
2019 ◽  
Vol 9 (4) ◽  
pp. 168
Author(s):  
Yorleydy Ruiz Moreno ◽  
Silvia Tavares Donato ◽  
Fátima Nogueira ◽  
Marcelo Sousa Silva
Keyword(s):  
Clinical History ◽  
Negative Control ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Response ◽  
Serum Samples ◽  
Serological Tests ◽  
Serological Reactivity ◽  
History Of ◽  
Low Levels ◽  
Antigenic Reactivity

Early diagnosis of malaria reduces disease, prevents deaths, and contributes to decreased malaria transmission. The use of specific and sensitive antigens in the execution of serological diagnostics may have an impact on the transmission of the disease. However, many individuals cannot be easily diagnosed by serological tests due to low levels of antibodies in the serum. Using two different Enzyme-Linked Immunosorbent Assay (ELISA) tests (a commercial and an in-house ELISA), a total of 365 serum samples from individuals with a clinical history of malaria were analyzed. From the serum samples analyzed, 192 (53%) samples from the commercial ELISA and 219 (60%) samples from the in-house ELISA presented positive serological reactivity to malaria. The concordance of the samples tested (n = 365) between both ELISAs was of 67% (n = 242), and with the negative control was 100% (n = 17). We demonstrated that the in-house ELISA showed high antigenic reactivity to Plasmodium falciparum antigens when compared with the commercial ELISA. The degree of concordance of both ELISAs suggested the possibility of existence of other P. falciparum antigens present in the crude extract of P. falciparum that are important in the serological response during malaria infection.

scholarly journals Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Harisankar Singha ◽  
Praveen Malik ◽  
Sachin K. Goyal ◽  
Sandip K. Khurana ◽  
Chiranjay Mukhopadhyay ◽  
...  
Keyword(s):  
Indirect Elisa ◽  
Expression System ◽  
Cross Reactivity ◽  
Amino Acid Sequences ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Diagnostic Efficacy ◽  
Diagnostic Potential ◽  
Prokaryotic Expression System ◽  
Control Serum

Objective. To express truncated TssB protein ofBurkholderia malleiand to evaluate its diagnostic efficacy for serological detection of glanders among equines.Materials and Methods. In an attempt to develop recombinant protein based enzyme-linked immunosorbent assay (ELISA), N-terminal 200 amino acid sequences ofB. malleiTssB protein—a type 6 secretory effector protein—were expressed in prokaryotic expression system. Diagnostic potential of recombinant TssB protein was evaluated in indirect ELISA using a panel of glanders positive (n=49), negative (n=30), and field serum samples (n=1811). Cross-reactivity of the assay was assessed with equine disease control serum and human melioidosis positive serum.Results. In comparison to CFT, diagnostic sensitivity and specificity of ELISA were 99.7% and 100%, respectively.Conclusions. The indirect ELISA method using the truncated TssB offered safer and more rapid and efficient means of serodiagnosis of glanders in equines. These data highlight the use of TssB as potential diagnostic antigen for serological diagnosis of glanders.

scholarly journals rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

AMB Express ◽  
2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Antonio Pedro Fróes de Farias ◽  
José Tadeu Raynal Rocha Filho ◽  
Silvana Beutinger Marchioro ◽  
Luan Santana Moreira ◽  
Andressa Souza Marques ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Recombinant Proteins ◽  
Corynebacterium Pseudotuberculosis ◽  
Semiarid Region ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Control Programs ◽  
Epidemiological Surveys ◽  
Purified Antigen

Abstract Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages, which include acceptable cost-effectiveness, applicability, sensitivity and specificity. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development due to its extracellular or on the cell surface location, which allows a better recognition by the immune system. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that ELISA-rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that ELISA-rSodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs. Trial registration AEC No 4958051018. 12/18/2018, retrospectively registered

scholarly journals Quantitative SARS-CoV-2 antibody screening of healthcare workers in the southern part of Kyoto city during the COVID-19 peri-pandemic period

2020 ◽  
Author(s):  
Kohei Fujita ◽  
Shinpei Kada ◽  
Osamu Kanai ◽  
Hiroaki Hata ◽  
Takao Odagaki ◽  
...  
Keyword(s):  
Healthcare Workers ◽  
Early Stage ◽  
Case Fatality ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igg Antibody ◽  
Kyoto City ◽  
History Of ◽  
Heavy Burden ◽  
Antibody Levels

Background: The coronavirus disease-2019 (COVID-19) pandemic is associated with a heavy burden on the mental and physical health of patients, regional healthcare resources, and global economic activity. While our understanding of the incidence and case-fatality rates increases, data on seroprevalence of antibodies against the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) in healthcare workers during the peri-pandemic period is insufficient. This study quantitatively evaluated seroprevalence of SARS-CoV-2 antibody in healthcare workers in the southern part of Kyoto city, Japan. Methods: We prospectively recruited healthcare workers from a single hospital between April 10 and April 20, 2020. We collected serum samples from these participants and quantitatively evaluated SARS-CoV-2 IgG antibody levels by enzyme-linked immunosorbent assay. Results: Five (5.4%), 15 (16.3%), and 72 (78.3%) participants showed positive, borderline, and negative serum SARS-CoV-2 IgG antibody status, respectively. We found the mean titer associated with each antibody status (overall, positive, borderline, and negative) was clearly differentiated. Participants working at the otolaryngology department and/or having a history of seasonal common cold symptoms had a significantly higher titer of SARS-CoV-2 IgG antibody (p=0.046, p=0.046, respectively). Conclusions: Five (5.4%) and 15 (16.3%) participants tested positive and borderline, respectively, for SARS-CoV-2 IgG antibody during the COVID-19 peri-pandemic period. These rates were higher than expected based on government situation reports. The present findings suggest that COVID-19 was already spread in the southern part of Kyoto city at the early stage of pandemic.

scholarly journals rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

2020 ◽  
Author(s):  
Antonio Pedro Fróes de Farias ◽  
José Tadeu Raynal Rocha Filho ◽  
Silvana Beutinger Marchioro ◽  
Luan Santana Moreira ◽  
Andressa Souza Marques ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Recombinant Proteins ◽  
Corynebacterium Pseudotuberculosis ◽  
Semiarid Region ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Control Programs ◽  
Epidemiological Surveys ◽  
Purified Antigen

Abstract Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that recombinant SodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.

scholarly journals Improved Serodiagnosis of Bovine Brucellosis by Novel Synthetic Oligosaccharide Antigens Representing the Capping M Epitope Elements of Brucella O-Polysaccharide

2015 ◽  
Vol 53 (4) ◽  
pp. 1204-1210 ◽  
Author(s):  
John McGiven ◽  
Laurence Howells ◽  
Lucy Duncombe ◽  
Judy Stack ◽  
N. Vijaya Ganesh ◽  
...  
Keyword(s):  
Brucella Melitensis ◽  
Effective Means ◽  
Cost Effective ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Content Type ◽  
Diagnostic Potential ◽  
Antibody Titers ◽  
Diagnostic Capabilities ◽  
Synthetic Antigens

Members of the genusBrucellahave cell wall characteristics of Gram-negative bacteria, which in the most significant species includes O-polysaccharide (OPS). Serology is the most cost-effective means of detecting brucellosis, as infection with smooth strains ofBrucellaleads to the induction of high antibody titers against the OPS, an unbranched homopolymer of 4,6-dideoxy-4-formamido-d-mannopyranosyl residues (d-Rha4NFo) that are variably α(1→2)- and α(1→3)-linked. Sixd-Rha4NFo homo-oligosaccharides were synthesized, each containing a single α(1→3) link but with a varied number of α(1→2) links. After conjugation to bovine serum albumin (BSA), glycoconjugates 1 to 6 were used to develop individual indirect enzyme-linked immunosorbent assays (iELISAs). The diagnostic capabilities of these antigens were applied to panels of cattle serum samples, including those falsely positive in conventional assays, and the results were compared with those of the complement fixation test (CFT), serum agglutination test (SAT), fluorescent polarization assay (FPA), smooth lipopolysaccharide (sLPS) iELISA, and competitive enzyme-linked immunosorbent assay (cELISA) methods. Results from field serum samples demonstrated that all of the synthetic antigens had excellent diagnostic capabilities. Assays developed with the α(1→3)-linked disaccharide conjugate 1 were the best at resolving false-positive serological results. This was supported by the results from serum samples derived from experimentally infected cattle. Data from synthetic trisaccharide antigens 2 and 3 and tetrasaccharide antigen 4 identified an OPS epitope equally common to allBrucella abortusandBrucella melitensisstrains but unique toBrucella. Synthetic oligosaccharide conjugates function as effective surrogates for naturally derived antigens. The creation of discrete OPS epitope antigens reveals not only the previously untapped diagnostic potential within this key diagnostic structure but also holds significance for the design of brucellosis vaccines and diagnostics that enable the differentiation of infected from vaccinated animals.

scholarly journals rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

2020 ◽  
Author(s):  
Antonio Pedro Fróes de Farias ◽  
José Tadeu Raynal Rocha Filho ◽  
Silvana Beutinger Marchioro ◽  
Luan Santana Moreira ◽  
Andressa Souza Marques ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Recombinant Proteins ◽  
Corynebacterium Pseudotuberculosis ◽  
Semiarid Region ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Control Programs ◽  
Epidemiological Surveys ◽  
Purified Antigen

Abstract Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that recombinant SodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.

scholarly journals rSodC is a potential antigen to diagnose Corynebacterium pseudotuberculosis by enzyme-linked immunoassay

2020 ◽  
Author(s):  
Antonio Pedro Fróes de Farias ◽  
José Tadeu Raynal Rocha Filho ◽  
Silvana Beutinger Marchioro ◽  
Luan Santana Moreira ◽  
Andressa Souza Marques ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Recombinant Proteins ◽  
Corynebacterium Pseudotuberculosis ◽  
Semiarid Region ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Control Programs ◽  
Epidemiological Surveys ◽  
Purified Antigen

Abstract Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages, which include acceptable cost-effectiveness, applicability, sensitivity and specificity. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development due to its extracellular or on the cell surface location, which allows a better recognition by the immune system. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that ELISA-rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that ELISA-rSodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.

scholarly journals rSodC is a Potential Antigen to Diagnose Corynebacterium Pseudotuberculosis by Enzyme-linked Immunoassay

2020 ◽  
Author(s):  
Antonio Pedro Fróes de Farias ◽  
José Tadeu Raynal Rocha Filho ◽  
Silvana Beutinger Marchioro ◽  
Luan Santana Moreira ◽  
Andressa Souza Marques ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Recombinant Proteins ◽  
Corynebacterium Pseudotuberculosis ◽  
Semiarid Region ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Serological Tests ◽  
Control Programs ◽  
Epidemiological Surveys ◽  
Purified Antigen

Abstract Caseous lymphadenitis (CL) is a chronic infectious disease that affects sheep and goats. Many serological tests have been developed to detect the disease; one of the most widely used is the enzyme-linked immunosorbent assay (ELISA), due to its advantages. ELISA formulations using recombinant proteins can exhibit significant sensitivity and specificity when using a single purified antigen. DTxR, Trx, TrxR, LexA, SodC, SpaC, NanH, and PknG recombinant proteins can be considered target proteins for ELISA development. Therefore, the objectives of this study were to evaluate the antigenic reactivity of Corynebacterium pseudotuberculosis recombinant proteins in goat and sheep serum. Of eight proteins evaluated, rSodC was selected for validation assays with small ruminant serum samples from the semiarid region of the state of Bahia, Brazil. Validation assays with goat serum samples showed that rSodC presented sensitivity and specificity of 96% and 94%, respectively. Validation assays with sheep serum showed that recombinant SodC exhibited sensitivity and specificity of 95% and 98%, respectively. Analysis of 756 field serum samples showed that rSodC identified 95 positive samples (23%) in goats and 75 positive samples (21%) in sheep. The ELISA with recombinant SodC protein developed in this study discriminated positive and negative serum samples with high levels of sensitivity and specificity. This formulation is promising for epidemiological surveys and CL control programs.

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