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Journal of Clinical Microbiology, August 1999, p. 2548-2552, Vol. 37, No. 8
0095-1137/99/$04.00+0

Molecular Epidemiological Study of Haemophilus influenzae Serotype b Strains Obtained from Children with Meningitis in Japan

Toshihiro Mitsuda,1,2,* Haruo Kuroki,3 Nobuyasu Ishikawa,3 Tomoyuki Imagawa,2 Schuichi Ito,2 Takako Miyamae,2 Masaaki Mori,2 Suzuko Uehara,3 and Shumpei Yokota2

Division of Clinical Laboratory Medicine1 and Department of Pediatrics,2 School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama City, 236-0004, and Department of Pediatrics, Chiba University School of Medicine, 1-8-1 Inohana, Chuoh-ku, Chiba City, Chiba, 260-0856,3 Japan

Received 19 August 1998/Returned for modification 22 November 1998/Accepted 21 May 1999

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.

* Corresponding author. Mailing address: Department of Pediatrics, School of Medicine, Yokohama City University, 3-9 Fukuura, Kanazawa-ku, Yokohama City, 236-0004, Japan. Phone: 81-45-787-2671. Fax: 81-45-784-3615. E-mail: tmitsuda{at}med.yokohama-cu.ac.jp.

Journal of Clinical Microbiology, August 1999, p. 2548-2552, Vol. 37, No. 8
0095-1137/99/$04.00+0


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