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.
Mitsuda
T
T
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
Kuroki
H
H
Ishikawa
N
N
Imagawa
T
T
Ito
S
S
Miyamae
T
T
Mori
M
M
Uehara
S
S
Yokota
S
S
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