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Sodium dodecyl sulphate-polyacrylamide gel electrophoresis of whole cell protein, immunoblotting with normal human serum and restriction endonuclease analysis using Taq I enzyme were applied to 38 clinically significant isolates of Moraxella (Branhamella) catarrhalis obtained during a suspected outbreak of nosocomial infection. Each of 18 strains had individual profiles by at least two of the three methods (unique strains). The remaining 20 strains were assigned to five groups (A-E) on the basis of similarity by at least two of the three methods. Isolates within groups A, D and E were homologous by all three methods. Immunoblot groups B and C had two distinct whole cell protein profiles (B1 and B2) but indistinguishable restriction endonuclease profiles (group B/C). This emphasizes the need to use more than one technique in characterizing strains from suspected outbreaks of nosocomial infection. Grouped strains were more likely to originate from the same hospital ward than unique strains and were associated with a significantly longer median time from patient admission to strain isolation (14 versus 3.5 days, p<0.005). Furthermore, the β-lactamase activity was homologous within the groups. The results suggest that nosocomial infection involving several distinct Moraxella catarrhalis strains persisted over a period of months, involving at least 20 patients on three different wards. Such infection is probably common in wards harbouring suitably predisposed patients. The mode of transmission remains to be elucidated, but the above three techniques possess sufficient reproducibility and discriminatory ability to constitute suitable investigative tools. [ABSTRACT FROM AUTHOR]