A generic mechanism in Neisseria meningitidis for enhanced resistance against bactericidal antibodies.

The presence of serum bactericidal antibodies is a proven correlate of protection against systemic infection with the important human pathogen Neisseria meningitidis. We have identified three serogroup C N. meningitidis (MenC) isolates recovered from patients with invasive meningococcal disease that resist killing by bactericidal antibodies induced by the MenC conjugate vaccine. None of the patients had received the vaccine, which has been successfully introduced in countries in North America and Europe. The increased resistance was not caused by changes either in lipopolysaccharide sialylation or acetylation of the alpha2-9-linked polysialic acid capsule. Instead, the resistance of the isolates resulted from the presence of an insertion sequence, IS1301, in the intergenic region (IGR) between the sia and ctr operons, which are necessary for capsule biosynthesis and export, respectively. The insertion sequence led to an increase in the transcript levels of surrounding genes and the amount of capsule expressed by the strains. The increased amount of capsule was associated with down-regulation of the alternative pathway of complement activation, providing a generic mechanism by which the bacterium protects itself against bactericidal antibodies. The strains with IS1301 in the IGR avoided complement-mediated lysis in the presence of bactericidal antibodies directed at the outer membrane protein, PorA, or raised against whole cells.

[Evaluation of xylose galactosidase (XG) culture medium for the isolation of enteropathogens]. / Evaluación del medio xilosa galactosidasa (XG) para el aislamiento de enteropatógenos.

INTRODUCTION: The chromogenic medium, XG, was evaluated and compared to conventional media for the isolation of Salmonella spp., Shigella spp., Yersinia enterocolitica and Aeromonas spp. METHODS: A total of 1226 human stool samples were inoculated on XG, MacConkey agar, Salmonella-Shigella agar (SS), selenite broth, blood-ampicillin agar and cefsulodin-Irgasan-novobiocin agar (CIN). RESULTS: The 235 positive cultures included the following: 229 Salmonella spp., 3 Shigella spp., 2 Yersinia enterocolitica and one Aeromonas spp. Among the 229 containing Salmonella spp., 100 were detected on both XG and conventional media and the 129 remaining were detected only on conventional media; recovery of Salmonella spp. on conventional media was significantly higher with respect to XG medium (p < 0.005). The 3 isolates of Shigella spp. were obtained on XG, the 2 isolates of Yersinia enterocolitica were recovered on CIN agar and the single isolate of Aeromonas spp. was obtained both on XG and blood-ampicillin agar. Colonies suspected to be some of the enteropathogens investigated were present in 791 of the negative stool samples. Among these false-positives 441 (35.9%) were obtained from XG, 142 (11.6%) after selenite enrichment, 132 (10.8%) from MacConkey agar and 76 (6.2%) from SS agar. Most of the false-positive isolates obtained on XG medium were consistent with Salmonella spp. (n = 408). CONCLUSIONS: XG chromogenic medium showed low sensitivity (64%) and specificity (69%) for the detection of Salmonella spp. Recovery of Shigella spp. on XG medium in three samples may have been due to the immediate processing of the samples. We conclude that XG chromogenic medium can not be recommended as an alternative to currently used conventional media.

Antigenic and/or phase variation of PorA protein in non-subtypable Neisseria meningitidis strains isolated in Spain.

The PorA protein is a potential candidate as a vaccine component against meningococcal disease. However, this protein experiences antigenic variation and is subject to phase variations to evade immune selective pressure. In this study, the mechanisms responsible for altered expression of the PorA protein were analysed in 50 non-subtypable strains isolated from patients with meningococcal disease in Spain. The porA gene was amplified from 47 of the 50 strains. The majority of isolates were not recognized by the subtyping panel, as a result of non-synonymous base changes in the variable regions of the porA gene. Two of these strains revealed a premature stop codon before the variable region VR1 of PorA due to a single base-pair substitution at position 109 of the porA coding region. Another two presented a homopolymeric tract of eight adenine residues in the coding region, producing a DNA strand-slippage mechanism and PorA phase variation.

Sequencing of Neisseria meningitidis penA gene: the key to success in defining penicillin G breakpoints.

Testing of susceptibility to penicillin G by E-test and sequencing of an internal fragment of the penA gene were done for 43 meningococcal strains. Those strains for which the MIC was >/=0.094 micro g/ml showed mosaic alleles, so 0.094 micro g/ml is suggested as the penicillin G intermediate breakpoint when E-test is used.