Separation of Haemophilus influenzae type b subtypes by numerical analysis

Abstract

Outer membrane proteins (OMPs) were prepared from 20 previously established subtypes of Haemophilus influenzae type b. Proteins were separated by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (11% acrylamide). Each lane in the gels contained two internal molecular weight standards. One central lane contained a range of molecular weight standards, which were used to calculate a molecular weight curve for each gel. Migration distances of the OMPs were determined with a soft laser-scanning densitometer, and the distances were normalized by using the mean migration distances of the internal standards. The protein patterns of all subtypes were compared by a recently described method (B.D. Plikaytis, G.M. Carlone, and B.B. Plikaytis, J. Gen. Microbiol. 132:2653-2660, 1986). All subtypes could be differentiated by this method. The ability to store and compare numerous OMP patterns from different isolates of H. influenzae type b, separated with a single homogeneous polyacrylamide gel, will facilitate the continued development of a subtyping system based on these proteins.