betadex and Whooping-Cough

Cyclodextrin solid medium (CSM) developed by us was evaluated to be a suitable synthetic medium for the clinical isolation of Bordetella pertussis when compared with Bordet-Gengou (BG) medium. The addition of 5 micrograms of cephalexin (CEX) per ml to CSM not only supported the good growth of B. pertussis but also sufficiently suppressed the growth of nasopharyngeal flora. During period 1 of this study, nasopharyngeal specimens from 60 patients with clinical pertussis were inoculated on CSM supplemented with 5 micrograms of CEX per ml. The isolation rate was 70% (42 of 60). To confirm the efficacy of CSM, another study was performed. During period 2 of this study, nasopharyngeal specimens were cultured on both CSM and BG medium, each with 5 micrograms of CEX per ml. The comparative isolation rates were 100% (40 of 40 specimens from 29 patients) on CSM with 5 micrograms of CEX and 65% (26 out of 40) on BG medium with 5 micrograms of CEX. The excellent efficacy of CSM as measured by the isolation rate was thought to be due to the poor nutrition of this medium for the growth of nasopharyngeal bacteria. CSM retained its efficacy in clinical isolations even after 3 months of storage in a refrigerator. These data led us to conclude that CSM with 5 micrograms of CEX was much better than BG medium with 5 micrograms of CEX, determined by both the isolation rate and preservativity considerations, and that CSM with 5 micrograms of CEX per ml can be successfully used instead of BG medium as a medium for the clinical isolation of B. pertussis.

Topics: beta-Cyclodextrins; Bordetella pertussis; Cephalexin; Child; Culture Media; Cyclodextrins; Dextrins; Humans; Nasopharynx; Starch; Whooping Cough

Heptakis (2,6-0 dimethyl)beta-cyclodextrin (MeCD) which permits the growth of single colonies of Bordetella pertussis Tohama phase I on Stainer-Scholte medium solidified with agar also enhanced the production of pertussis toxin (PT). More than one hundred times the amount of PT was produced in Stainer-Scholte medium with MeCD in shake culture than was produced in MeCD-free medium. A maximum of 50 mg PT protein was produced per liter of culture broth as estimated by in vitro and in vivo assays. The production of filamentous hemagglutinin (FHA) was several hundred times greater when B. pertussis was grown in shake cultures with MeCD than when growth was in MeCD-free shake cultures. The FHA content of the production medium was confirmed by enzyme-linked immunosorbent assays and electron microscopy. Evaluation of an acellular vaccine containing PT and FHA detoxified with formaldehyde showed that it was protective in the intracerebral challenge mouse potency assay.

Topics: Animals; beta-Cyclodextrins; Bordetella pertussis; Cell Division; Cyclodextrins; Dextrins; Hemagglutinins; Mice; Pertussis Toxin; Pertussis Vaccine; Starch; Virulence Factors, Bordetella; Whooping Cough