scholarly journals Evaluation of a Western Blot Method for the Detection of Yersinia Antibodies: Evidence of Serological Cross-Reactivity between Yersinia Outer Membrane Proteins and Borrelia burgdorferi

2005 ◽  
Vol 12 (11) ◽  
pp. 1269-1274 ◽  
Author(s):  
Mindy L. Rawlins ◽  
Cecilia Gerstner ◽  
Harry R. Hill ◽  
Christine M. Litwin
Keyword(s):  
Membrane Proteins ◽  
Borrelia Burgdorferi ◽  
Western Blot ◽  
Outer Membrane ◽  
Sensitivity And Specificity ◽  
Reactive Arthritis ◽  
Outer Membrane Proteins ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Western Blot Assay

ABSTRACT Yersinia enterocolitica and Yersinia pseudotuberculosis have been identified as causative organisms of reactive arthritis in humans. We evaluated a Western blot assay which uses Yersinia outer membrane proteins as antigens for the detection of Yersinia antibodies as a replacement for the complement fixation (CF) assay. Clinical agreement, sensitivity, and specificity were determined by testing 19 positive and 21 negative serum samples by the CF assay, Western blot assay, and enzyme-linked immunosorbent assay (ELISA). The CF assay and ELISA were compared to the Western blot assay, which was the reference method used in this study. Sera with antibodies that could potentially cross-react with Yersinia were also tested by the Western blot assay. The agreement, sensitivity, and specificity of the CF method were 61%, 26%, and 95%, respectively; and those for the ELISA were 89%, 95%, and 82%, respectively. The prevalences of Yersinia antibodies in 50 healthy donors were 6% for immunoglobulin G (IgG), 2% for IgA, and 2% for IgM. Sera positive for Bartonella henselae, Brucella, Chlamydia pneumoniae, and Rickettsia rickettsii antibodies showed cross-reactivity by the Western blot assay. The highest cross-reactivity was observed with Borrelia burgdorferi; 5 of 11 (45%) specimens were cross-reactive by the IgM-specific assay. Overall, the Western blot assay performs acceptably and is more sensitive than the CF assay, warranting replacement of the CF assay in the laboratory. Due to the evidence of cross-reactivity, particularly with B. burgdorferi, which can cause an oligoarthritis similar to reactive arthritis, the diagnosis of reactive arthritis should be based on clinical findings and complete serologic analysis of the potential causative infectious pathogens.

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.

scholarly journals Protein and antigen profiles of Leptospira interrogans serovar Hardjo

2009 ◽  
Vol 39 (9) ◽  
pp. 2539-2543
Author(s):  
Bárbara Nobre Lafetá ◽  
Elaine Cristina de Castro ◽  
Nivaldo da Silva
Keyword(s):  
Immune Response ◽  
Membrane Proteins ◽  
Western Blot ◽  
Outer Membrane ◽  
Protein Profile ◽  
Outer Membrane Proteins ◽  
Rabbit Antiserum ◽  
Leptospira Interrogans ◽  
Sds Page ◽  
Hyperimmune Rabbit

The protein profile of the outer membrane of Leptospira interrogans serovar Hardjo subtype hardjoprajitno associated with the bovine natural immune response was investigated. The outer membrane proteins were extracted utilizing Triton X114 and precipitated with acetone. The protein sample was then resolved by SDS-PAGE and reacted in western blot against sera from a hyperimmune rabbit and from naturally infected bovines. In silver stained gels, 14 protein bands were observed, among which four proteins, with 22, 29, 47 and 63kDa, appeared as major constituents. Western blot tests with hyperimmune rabbit antiserum detected bands corresponding to proteins with 35; 27; 24; 21; 17 and 14kDa, while 32kDa and 45kDa proteins were the most immunoreactive with sera from naturally infected bovines.

scholarly journals Serum Antibody Responses in Ethiopian Meningitis Patients Infected with Neisseria meningitidis Serogroup A Sequence Type 7

2007 ◽  
Vol 14 (4) ◽  
pp. 451-463 ◽  
Author(s):  
Gunnstein Norheim ◽  
Abraham Aseffa ◽  
Mohammed Ahmed Yassin ◽  
Getahun Mengistu ◽  
Afework Kassu ◽  
...  
Keyword(s):  
Membrane Proteins ◽  
Outer Membrane ◽  
Acute Phase ◽  
Outer Membrane Proteins ◽  
Serum Antibody ◽  
Outer Membrane Vesicles ◽  
Sequence Type ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Complement ◽  
Convalescent Phase

ABSTRACT To elucidate critical components of protective immune responses induced during the natural course of serogroup A meningococcal disease, we studied acute-, early-convalescent-, and late-convalescent-phase sera from Ethiopian patients during outbreaks in 2002 to 2003. Sera were obtained from laboratory-confirmed patients positive for serogroup A sequence type 7 (ST-7) meningococci (A:4/21:P1.20,9) (n = 71) and from Ethiopian controls (n = 113). The sera were analyzed using an enzyme-linked immunosorbent assay to measure levels of immunoglobulin G (IgG) against serogroup A polysaccharide (APS) and outer membrane vesicles (OMVs) and for serum bactericidal activity (SBA) using both rabbit and human complement sources. Despite relatively high SBA titers and high levels of IgG against APS and OMVs in acute-phase patient sera, significant increases were seen in the early convalescent phase. Antibody concentrations returned to acute-phase levels in the late convalescent phase. Considering all patients' sera, a significant but low correlation (r = 0.46) was observed between SBA with rabbit complement (rSBA) using an ST-5 reference strain and SBA with human complement (hSBA) using an ST-7 strain from Ethiopia. While rSBA demonstrated a significant linear relation with IgG against APS, hSBA demonstrated significant linear relationships with IgG against both APS and OMV. This study indicates that antibodies against both outer membrane proteins and APS may be important in providing the protection induced during disease, as measured by hSBA. Therefore, outer membrane proteins could also have a role as components of future meningococcal vaccines for the African meningitis belt.

scholarly journals Development of a Serological Test forHaemophilus ducreyi for Seroprevalence Studies

2000 ◽  
Vol 38 (4) ◽  
pp. 1520-1526 ◽  
Author(s):  
Christopher Elkins ◽  
Kyungcheol Yi ◽  
Bonnie Olsen ◽  
Christopher Thomas ◽  
Kevin Thomas ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Proteins ◽  
Enzyme Immunoassay ◽  
Outer Membrane ◽  
Sensitivity And Specificity ◽  
Blood Donors ◽  
Serological Test ◽  
Outer Membrane Proteins ◽  
Haemophilus Ducreyi ◽  
Genital Ulcers

We developed a new enzyme immunoassay (rpEIA) for use in determining the seroprevalence of chancroid. Three highly conserved outer membrane proteins from Haemophilus ducreyi strain 35000 were cloned, overexpressed, and purified from Escherichia coli for use as antigens in the rpEIA. Serum specimens from patients with and without chancroid were assayed to determine optimum sensitivity and specificity and to establish cutoff values. On the basis of these data, rpEIA was found to be both sensitive and specific when used to test a variety of serum specimens from patients with genital ulcers and urethritis and from healthy blood donors.

Sign in / Sign up

Export Citation Format

Share Document