Jpn. J. Infect. Dis., 55, 2002

Laboratory and Epidemiology Communications

Molecular Epidemiology of Group A Streptococci T Serotype 1
Daisuke Tanaka1,5*, Gyobu Yotaku1, Junko Isobe1, Shiho Hosorogi1, Miwako Shimizu1,
Koji Katori1, Yoshiyuki Nagai1, Yuichi Fuchi2, Jun Yatsuyanagi3, Shioko Saito3,
Jun Kudaka4 and Shinichi Nakamura5
1

Department of Bacteriology, Toyama Institute of Health,
Nakataikoyama 17-1, Kosugi, Toyama 939-0363,
2
Department of Bacteriology, Oita Prefectural Institute of Health and Environment,
Hogawaradai 2-51, Oita 870-0948,
3
Department of Microbiology, Akita Prefectural Institute of Public Health,Sensyu 6-6, Kubota-machi, Akita 010-0874,
4Department of Biological Sciences, Okinawa Prefectural Institute of Health and Environment,
Ozato 2085, Ozato-son, Okinawa 901-1202 and5
Department of Bacteriology, Graduate School of Medical Science, Kanazawa University,
Takara-machi 13-1, Kanazawa 920-8640
Communicated by Yutaka Miyazaki(Accepted July 4, 2002)

1990 in Akita and until 1992 in Toyama and Oita. The second
prevalent strains, pattern B, were isolated in these three areas
since 1990 but not before. All strains isolated in Okinawa
Prefecture from 1991 belonged to pattern B. Among pattern
A strains, either pattern A2 or A7 or both were dominantdepending upon the area. Pattern B1 was dominant among
pattern B strains in all the areas. Murase et al. (5) reported
that PFGE patterns of T1/M1 strains isolated from patientswith pharyngitis in Kanagawa Prefecture changed over 19881989, and that almost all strains showed dominant PFGE
patterns from 1989 possessed the speA gene. All these findings
indicate that the speA-positive clone of serotype T1/M1 GASstrains spread in Japan during 1989-1990 and replaced thespeA-negative clone in various areas of Japan.

Group A streptococcus (GAS) serotype T1/M1 has beencommonly associated with streptococcal infections in Japan(1). Pulsed-field gel electrophoresis (PFGE) analysis of T1
isolates obtained from patients in Toyama Prefecture revealed
that the PFGE pattern in 1987-1988 differed from that in 19911993 (2). Clonal changes of M1 strains on the basis of PFGE
were also observed in the United States during the late 1980s(3) and in New Zealand during the early 1980s (4). In thepresent study, we examined the PFGE patterns of T1 GAS
strains from four geographically distant areas in Japan during
1983-1998, with a view to clarify the communicability of theorganisms.
A total of 308 T1 GAS isolates from patients withoutcomplications were examined. SmaI PFGE analysis wasperformed by using CHEF-DRII (Bio-Rad Laboratories,Hercules, Calif., USA). Southern hybridization analysis wasperformed employing the Gene Images Random-PrimeLabeling and Detection System (Amersham Pharmacia Biotech,Buckinghamshire, UK) using probes of PCR amplified speA
(393 bp), speB (1,113 bp), and speC (540 bp).
Fourteen different PFGE patterns were found (Fig. 1). A
computer-generated dendrogram revealed two large clusters,
A and B (data not shown). Southern hybridization analysis
also showed the clear difference between patterns A and B
isolates (Fig. 2). The speA probe hybridized with none of theDNA fragments of the pattern A isolates but with a ~110 kb
fragment of all the pattern B isolates. The speB probehybridized with a ~160 kb fragment of all the pattern A
isolates and a ~200 kb fragment of all the pattern B isolates
except for pattern B3 isolate that showed a slightly larger
hybridizing band. The speC probe hybridized with a DNA
fragment of patterns A3, A7, and B2. Table 1 shows a common
tendency in the three distant areas, Akita, Toyama, and Oita
Prefectures, in which pattern A strains predominant before
1990 were gradually replaced by pattern B strains in 3 years
from 1990 to 1992. The pattern A strains were present until

Fig. 1. PFGE patterns of SmaI-digested chromosomal DNA of T1 group
A streptococci. M: λDNA ladder, Lane 1: PFGE pattern A1, Lane 2:
A2, Lane 3: A3, Lane 4: A4, Lane 5: A5, Lane 6: A6, Lane 7: A7,Lane 8: B1, Lane 9: B2, Lane 10: B3, Lane 11: B4, Lane 12: B5,Lane 13: B6, Lane 14: B7.

*Corresponding author: Tel: +81-766-56-5506, Fax: +81-766-567326
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Jpn. J. Infect. Dis., 55, 2002
Table 1. Distribution of Group A Streptococci T serotype 1
Place
Akita

ToyamaOita

Okinawa
Total

PFGE No. of1983-1989 1990 1991 1992 1993-1998
pattern isolates
A7
B1
B61820313

A1A2A3A4A7B1B2B3B4B7107114175
1112954
14A2A4A5A6A7B1B5211219392121111B1B4241308

51651962138128352
1112977214234

36110415153835104

21
2

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Fig. 2. Southern hybridization analysis of SmaI-digested DNA resolvedby PFGE. The same membrane was stripped and re-probed with thefollowing probes: speA (a), speB (b), and speC (c).

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