Listeria monocytogenes serotype 4b is a food-borne pathogen of public health concern, since it accounts for approximately 40% of human listeriosis cases. We have recently identified IspC, a surface-localized peptidoglycan hydrolase, as the antigen recognized by a number of monoclonal antibodies (MAbs) produced against a serotype 4b strain for diagnostic applications. To determine whether IspC, which is well conserved among various serotype 4b strains, is a useful diagnostic marker in antibody-based methods, we assessed the expression of IspC in L. monocytogenes cultured under normal and stress conditions. A functional promoter directing the transcription of the ispC gene was identified upstream of the ispC open reading frame by constructing a promoterless lacZ gene fusion with the putative ispC promoter region and by 5= rapid amplification of cDNA ends analysis. Using both the lacZ reporter gene system and immunofluorescent staining with an IspC-specific MAb, we provide evidence that IspC is expressed on the cell surface in all growth conditions tested (temperature, osmotic stress, pH, ethanol, oxidative stress, anaerobic conditions, carbon source, and type of growth media) that allow for cellular division, although the level of ispC gene expression varies. These results demonstrated the usefulness of IspC as an excellent diagnostic marker for the serotype 4b strains and imply that IspC, in conjunction with specific MAbs, can be targeted for detection and isolation of L. monocytogenes serotype 4b strains directly from food, environmental, and clinical samples with minimal or no need for culture enrichment.
Listeria monocytogenes is a Gram-positive pathogenic bacterium that can lead to listeriosis, a life-threatening opportunistic infection caused by the ingestion of contaminated foods. Clinical outcomes of listeriosis range from asymptomatic infection to nonspecific flu-like symptoms, gastroenteritis, septicemia, meningitis, and fetal infection followed by abortion in pregnant women (24). In the environment, L. monocytogenes is extremely hardy and actively divides between 3 and 45°C (26), in up to 10% salt (16), at a pH of between 4.4 and 9.2 (5), and under anaerobic conditions (15). The ability of L. monocytogenes to grow in extreme environments makes it a concern for the food industry, particularly in food-processing plants where ready-to-eat foods are prepared.L. monocytogenes is divided into 13 serotypes; however, 98% of human illness is caused by serotype 1/2a, 1/2b, and 4b strains (8). Serotype 4b strains account for more cases of human listeriosis than serotype 1/2a and 1/2b isolates combined, although 1/2a and 1/2b strains are much more commonly found in foods and the environment (24,25). This suggests that serotype 4b strains are specifically adapted to infecting human hosts (24, 25). Serotype 4b strains are also more often isolated from patients with meningoencephalitis than from patients where the infection has been limited to the bloodstream (24). Listeriosis patients also suffer a 26% mortality rate...