Molecular epidemiological study of Haemophilus influenzae serotype b strains obtained from children with meningitis in Japan.
We report an epidemiological study of 30 Haemophilus influenzae serotype b (Hib) strains derived from the cerebrospinal fluid of children with meningitis. The Hib strains were biotyped, tested for beta-lactamase production, and genotyped by long PCR-ribotyping, random amplified polymorphic DNA (RAPD) analysis, and genomic DNA restriction fragment length polymorphism (RFLP) analysis by pulsed-field gel electrophoresis (PFGE). The phenotypic study characterized 22 of the strains (73%) as biotype I. A genotypic study using long PCR-ribotyping with HaeIII restriction digestion showed no polymorphisms among these 30 Hib strains, but RAPD analysis with two sets of primers demonstrated two distinctive subtypes: one typical of the strains of biotype group II and the second characteristic of the strains of biotype groups I and IV. Each RAPD group was subtyped into several genotypic groups by PFGE-RFLP with SmaI digestion. The genotyping of clinically isolated Hib strains may help to elucidate transmission routes in community infections, endemicity, and the reasons for vaccine failure.
Division of Clinical Laboratory Medicine, School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama City, 236-0004, Japan. tmitsuda@med.yokohama-cu.ac.jp
T
T
Mitsuda
H
H
Kuroki
N
N
Ishikawa
T
T
Imagawa
S
S
Ito
T
T
Miyamae
M
M
Mori
S
S
Uehara
S
S
Yokota
D002675:Child, Preschool; D016521:Electrophoresis, Gel, Pulsed-Field; D016679:Genome, Viral; D006193:Haemophilus influenzae / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D007223:Infant; D007564:Japan / Q000453:epidemiology; D008581:Meningitis / Q000453:epidemiology / Q000382:microbiology*; D016133:Polymerase Chain Reaction / Q000379:methods; D012150:Polymorphism, Restriction Fragment Length; D016036:Seroepidemiologic Studies; D012698:Serologic Tests