Correlation between serological and sequencing analyses of the PorB outer membrane protein in the Neisseria meningitidis serotyping system.
The current serological typing scheme for Neisseria meningitidis is not comprehensive; a proportion of isolates are not serotypeable. DNA sequence analysis and predicted amino acid sequences were used to characterize the structures of variable-region (VR) epitopes on N. meningitidis PorB proteins (PorB VR typing). Twenty-six porB gene sequences were obtained from GenBank and aligned with 41 new sequences. Primary amino acid structures predicted from those genes were grouped into 30 VR families of related variants that displayed at least 60% similarity. We correlated VR families with monoclonal antibody (MAb) reactivities, establishing a relationship between VR families and epitope locations for 15 serotype-defining MAbs. The current panel of serotype-defining MAbs underestimates by at least 50% the PorB VR variability because reagents for several major VR families are lacking or because a number of VR variants within some families are not recognized by serotype-defining MAbs. These difficulties, also reported for serosubtyping based on the PorA protein, are shown as inconsistent results between serological and sequence analyses, leading to inaccurate strain identification and incomplete epidemiological data. The information from this study enabled the expansion of the panel of MAbs currently available for serotyping, by including MAbs of previously undetermined specificities. Use of the expanded serotype panel enabled us to improve the sensitivity of serotyping by resolving a number of formerly nonserotypeable strains. In most cases, this information can be used to predict the VR family placement of unknown PorB proteins without sequencing the entire porB gene. PorB VR typing complements serotyping, and a combination of both techniques may be used for full characterization of meningococcal strains. The present work represents the most complete and integrated data set of PorB VR sequences and MAb reactivities of serogroup B and C meningococci produced to date.
Sacchi
C T
CT
Bacteriology Division, Adolfo Lutz Institute, São Paulo, Brazil. cls9@cdc.gov
Lemos
A P
AP
Whitney
A M
AM
Solari
C A
CA
Brandt
M E
ME
Melles
C E
CE
Frasch
C E
CE
Mayer
L W
LW
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000145:classification* / Q000235:genetics* / Q000276:immunology; D015373:Bacterial Typing Techniques; D001483:Base Sequence; D000939:Epitopes; D006801:Humans; D007135:Immunoglobulin Variable Region; D008969:Molecular Sequence Data; D009345:Neisseria meningitidis / Q000145:classification* / Q000235:genetics / Q000276:immunology; D018272:Porins; D017422:Sequence Analysis, DNA; D012703:Serotyping; D013045:Species Specificity
D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D007135:Immunoglobulin Variable Region; D018272:Porins; C057699:porin protein, Neisseria