생물형 및 OMP P2 양상에 의한 국내 분리 Haemophilus infuenzae의 항원학적 분석

Abstract

To understand major antigenic protein profile and to analyze Onter membrane protein (OMP) P2 subtypes, 150 isolates including 14 Haemophilus influenzae serotype b (Hib) and, 136 of nontypeable H. influenzae (NTHI) strains were characterized by biotyping, SDS-PAGE, and immunoblotting with monoclonal antibodies. In NTHI biotyping, biotype I and II were the most representative ones, which includes 45 isolates (27.6%) and 31 isolates (19.0%), respectively. Hib strains were classified into biotype I, II, V, VI, and VIII. Any correlation could not be confirmed among the biotypes and the sources of specimen. Based on the SDS-PAGE patterns of OMP P2 protein, ranging from MW 32 KDa to 42 KDa, NTHI strains were classified into 8 subtypes. All of the serotype b strains showed identical pattern with the 38 KDa, except J18 which showed 36 KDa, OMP P2 protein profile. In SDS-PAGE, Hib OMP P2 showed distinctive difference from NTHI strains. Two monoclonal antibodies, 3F8 and 5E5, specific to the protein P2 were used to type the strains. Mab 5E5, which showed Haemophilus genus specificity, reacted with H. parainfluenzae 7901 and biotype aegyptius 11116, but not with N. meningitidis, S. pneumoniae, or E. coli. It was confirmed that the Mab 3F8 and 5E5 were directed against the surface exposed OMP epitope of immunized strain by immunogold electron microscopy. In conclusion, OMP P2 was a major antigenic protein, and was supposed to play an important role in antigen detection and to be used for the molecular epidemiologic study of H. influenzae isolates.