Development of aChlamydia suis-specific antibody enzyme-linked immunosorbent assay based on the use of a B-cell epitope of the polymorphic membrane protein C

ABSTRACT Recently, a novel 12-mer B-cell epitope, MKNNPIYSEGSL, in the type-specific region of equine herpesvirus 1 (EHV-1) glycoprotein G (gG) was identified and used as an antigen for enzyme-linked immunosorbent assay (Maeda et al., J. Clin. Microbiol. 42:1095-1098, 2004). Although our prototype strain, TH20p, possesses two repeat sequences containing the B-cell epitope, the EHV-4 NS80567 strain has two repeat sequences that are not identical. One repeat sequence stretch contained the B-cell epitope, while the other contained the 11-mer, MKNNP V YSESL (underlining indicates a different amino acid). In this study, heterogeneity of the type-specific region was compared among Japanese EHV-4 isolates. The 11-mer peptide, MKNNP V YSESL, specifically reacted with sera from horses naturally infected with EHV-4 but not with sera from horses experimentally infected with EHV-4 TH20p. The 11-mer peptide may be another B-cell epitope in the type-specific region.

Download Full-text

Identification of human linear B-cell epitope sites on the envelope glycoproteins of Crimean-Congo haemorrhagic fever virus

Epidemiology and Infection ◽

10.1017/s0950268814002271 ◽

2014 ◽

Vol 143 (7) ◽

pp. 1451-1456 ◽

Cited By ~ 4

Author(s):

D. GOEDHALS ◽

J. T. PAWESKA ◽

F. J. BURT

Keyword(s):

B Cell ◽

Cell Epitope ◽

Fever Virus ◽

Enzyme Linked Immunosorbent Assay ◽

Haemorrhagic Fever ◽

Serum Samples ◽

B Cell Epitope ◽

Crimean Congo Haemorrhagic Fever ◽

Peptide G ◽

Linear B

SUMMARYA peptide library was used to screen for regions containing potential linear B-cell epitope sites in the glycoproteins and nucleoprotein of Crimean-Congo haemorrhagic fever virus (CCHFV) in an enzyme-linked immunosorbent assay (ELISA). The library consisted of 156 peptides, spanning the nucleoprotein and mature GN and GC proteins in a 19-mer with 9-mer overlap format. Using pooled serum samples from convalescent patients to screen the library, six peptides were identified as potential epitope sites. Further testing of these six peptides with individual patient sera identified two of these peptides as probable epitope sites, with peptide G1451–1469 reacting to 13/15 and peptide G1613–1631 to 14/15 human sera. These peptides are situated on the GC protein at amino acid positions 1451–1469 (relative to CCHFV isolate SPU103/97) (TCTGCYACSSGISCKVRIH) and 1613–1631 (FMFGWRILFCFKCCRRTRG). Identified peptides may have application in ELISA for diagnostic or serosurveillance purposes.

Download Full-text

A linear B-cell epitope on the class 3 outer-membrane protein of Neisseria meningitidis recognized after vaccination with the Norwegian group B outer-membrane vesicle vaccine

Microbiology ◽

10.1099/13500872-141-7-1593 ◽

1995 ◽

Vol 141 (7) ◽

pp. 1593-1600 ◽

Cited By ~ 8

Author(s):

A. A. Delvig ◽

E. Wedege ◽

D. A. Caugant ◽

R. Dalseg ◽

J. Kolberg ◽

...

Keyword(s):

Membrane Protein ◽

B Cell ◽

Neisseria Meningitidis ◽

Outer Membrane ◽

Outer Membrane Protein ◽

Membrane Vesicle ◽

Cell Epitope ◽

Outer Membrane Vesicle ◽

B Cell Epitope ◽

Group B

Download Full-text

Competitive Enzyme-Linked Immunosorbent Assay Based on a Rhoptry-Associated Protein 1 Epitope Specifically Identifies Babesia bovis-Infected Cattle

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.10.1.38-43.2003 ◽

2003 ◽

Vol 10 (1) ◽

pp. 38-43 ◽

Cited By ~ 22

Author(s):

Will L. Goff ◽

Terry F. McElwain ◽

Carlos E. Suarez ◽

Wendell C. Johnson ◽

Wendy C. Brown ◽

...

Keyword(s):

Immunosorbent Assay ◽

Disease Surveillance ◽

Cell Epitope ◽

Babesia Bovis ◽

Enzyme Linked Immunosorbent Assay ◽

Specific Monoclonal Antibody ◽

C Terminus ◽

B Cell Epitope ◽

High Throughput Method ◽

Species Specific

ABSTRACT The competitive enzyme-linked immunosorbent assay (cELISA) format has proven to be an accurate, reliable, easily standardized, and high-throughput method for detecting hemoparasite infections. In the present study, a species-specific, broadly conserved, and tandemly repeated B-cell epitope within the C terminus of the rhoptry-associated protein 1 of the hemoparasite Babesia bovis was cloned and expressed as a histidine-tagged thioredoxin fusion peptide and used as antigen in a cELISA. The assay was optimized with defined negative and positive bovine sera, where positive sera inhibited the binding of the epitope-specific monoclonal antibody BABB75A4. The cELISA accurately differentiated animals with B. bovis-specific antibodies from uninfected animals and from animals with antibodies against other tick-borne hemoparasites (98.7% specificity). In addition, B. bovis-specific sera from Australia, Argentina, Bolivia, Puerto Rico, and Morocco inhibited the binding of BABB75A4, confirming conservation of the epitope. The assay first detected experimentally infected animals between 13 and 17 days postinfection, and with sera from naturally infected carrier cattle, was comparable to indirect immunofluorescence (98.3% concordance). The assay appears to have the characteristics necessary for an epidemiologic and disease surveillance tool.

Download Full-text