Antigen-specific serotyping of Neisseria gonorrhoeae: characterization based upon principal outer membrane protein

Abstract

Principal outer membrane protein (protein I) of Neisseria gonorrhoeae was prepared nearly free of lipopolysaccharide (LPS) and substantially purified from other membrane proteins by chromatography of partially purified gonococcal outer membranes over Sepharose 6B in the presence of deoxycholate at pH 9.0. This protein I of nine separate antigenic types was coated to polystyrene tubes and used in the enzyme-linked immunosorbent assay (ELISA) to measure antibody to protein I or in inhibition tests to quantitate protein I antigen. No significant inhibition of the ELISA test was produced by purified LPS from the strain used to prepare each of the protein I types or by whole gonococci bearing the same LPS but different protein I antigens as the strain used to produce a given protein I antigen. Of 125 strains of gonococci used as whole organisms to inhibit the protein I ELISA, 124 (99%) typed with one or more of the nine protein I types, and 35% of these typed with a single protein I serotype. Sixty-one of 65 (94%) strains from Seattle and Atlanta patients with disseminated gonococcal infection contained protein I serotype 1, and 16 of 24 (64%) strains from Seattle patients with salpingitis bore one or both of protein I serotypes 1 and 2.

Differentiation of outer membrane proteins from Salmonellaenterica serotypes using Fourier transform infrared spectroscopy and chemometrics

“…The migrational OMP pattern analysis using SDS-PAGE has been investigated on the gel after staining and destaining steps for sub-grouping bacterial serotypes (Barenkamp et al 1981;Buchanan and Hildebrandt 1981). However this conventional method often provides poor classification between bacterial isolates, especially for Salmonella serotypes, due to their high degree of similarity in major OMP patterns.…”

“…Outer membrane proteins (OMPs) of pathogenic bacteria exhibit immunogenic and virulent properties and have been investigated for diagnostic application of monoclonal or polyclonal antibodies (Yancey et al 1979;Hale et al 1983;Udhayakumar and Muthukuruppan 1987;Kerr et al 1992;Charles et al 1994). OMP profiles of many bacterial species have been characterized using SDS-PAGE (Munford et al 1975; Barenkamp et al 1981;Buchanan and Hildebrandt 1981;Achtman et al 1983). However, OMP pattern analysis of Salmonella species revealed a high level of homogeneity between various serotypes, and therefore using SDS-PAGE for bacterial differentiation can be problematic (Helmuth et al 1985;Malik et al 1999).…”

Construction of isogenic gonococci with variable porin structure: effects on susceptibility to human serum and antibiotics

“…There are significant structural and chemical differences between proteins of the two serotypes, as demonstrated by their size and susceptibility to cleavage by various proteolytic enzymes (Blake efa/., 1981). There is some correlation between PI serotype and the clinical state of the disease, since disseminated gonococcal infections are usually caused by PIA strains, while PIB strains are more commonly isolated from local infections of the mucosal surfaces (Buchanan and Hildebrandt, 1981). PI serotype has also been associated witti traits such as antibiotic resistance (Bygdeman et at., 1984;Danielsson et at., 1986), serum resistance (Cannon etat., 1981;Hitdebrandt ef ai, 1978), auxotype (Brunham er a/., 1985).…”

Outer membrane proteins of Brucella abortus: isolation and characterization

“…Although the proteins under investigation are structural constituents of the cell, and therefore unlikely to function primarily as virulence factors, they may induce immune responses useful in diagnosis and in conferring protective immunity. Outer membrane proteins have been used in other genera in serological classification (7,8,18,24) and as vaccines (1, 13,31). Because B. abortus is a facultative intracellular parasite devoid of capsule, flagella, and pili, it seemed reasonable that protective immunity would include a cell-mediated immune response to one or more protein constituents of the cell wall.…”