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Journal of Clinical Microbiology, December 2001, p. 4526-4528, Vol. 39, No. 12
Service de Microbiologie, Hôpital
Robert Debré, 75019 Paris,1 and
Service de Bactériologie, Hôpital Saint Vincent
de Paul, 75014 Paris,2 France
Received 9 April 2001/Returned for modification 22 June
2001/Accepted 21 August 2001
We examined the genetic diversity of serotype V group B
streptococcus (GBS) isolates in the Paris area and compared them with the predominant American serotype V clone. Pulsed-field gel
electrophoresis yielded 11 patterns for 64 French GBS. One pattern was
obtained with 60% of the isolates tested and was indistinguishable
from that of the predominant American clone.
Group B streptococci (GBS)
are the main cause of severe infection in both infants and adults
(2, 20, 21). GBS are serotyped on the basis of their
capsular polysaccharide, of which nine different serotypes have been
described (10, 15). The classical serotypes Ia, Ib, II,
and III are the predominant cause of disease in neonates. Recently,
serotype V GBS have emerged as a new cause of GBS infection or
colonization in children and adults (13, 18, 19). Indeed, population-based surveillance of GBS in the 1990s indicated that serotype V was responsible for 10 to 15% of neonatal GBS infections in
the United States (7, 13, 17) and was the most common serotype isolated from nonpregnant adults with invasive disease (13). Moreover, data from the Centers for Disease Control
and Prevention (Atlanta, Ga.) have shown the emergence of a serotype V
clonal type (8).
In France, too, recent studies point to a significant shift in the
distribution of GBS serotypes, with serotype V emerging as one of the
major serotypes recovered from neonates (1, 11, 16).
We examined the genetic diversity of French serotype V GBS clinical
isolates in the Paris area and compared them with the predominant
American clone.
We studied a collection of 64 serotype V GBS isolates recovered between
January 1998 and January 2000 in the Paris area. The isolates were
obtained from genital specimens from pregnant women (n = 30) or from gastric fluid or ear specimens from colonized or infected
newborns (n = 34). GBS isolates were confirmed at the
species level by standard laboratory methods and were serotyped with a
commercial latex agglutination kit (Streptex; Murex Diagnostics UK). The isolates were stored in Todd-Hewitt broth with 20%
glycerol at Pulsed-field gel electrophoresis (PFGE) of serotype V GBS strains was
performed as previously described (12). SmaI
restriction enzyme chromosomal digests were separated with a
Bio-Rad contour-clamped homogenous electric field mapper with a
switch time of 0.85 to 35.38 s for 22 h and 35 min at a 120°
angle with a voltage gradient of 6 V/cm at 14°C. The DNA size
standard was a lambda DNA ladder (Bio-Rad). The gels were stained with
ethidium bromide and photographed under UV light. PFGE banding patterns
were compared visually. Strains were considered genetically
distinguishable if their restriction patterns differed by three or more
bands (24). Banding patterns were also compared using a
computer system (Biocapt; Vilber-Lourmat) and whole-band analyzer
software (Biogène; Vilber-Lourmat). Cluster analysis (unweighted
pair group average) was used to calculate similarity and dissimilarity
among GBS isolates. A difference was considered significant if the
similarity coefficient was <80%. The results were compared with those
obtained with the predominant American clonal serotype V GBS strain
(8).
PFGE typing of the 64 French serotype V GBS isolates yielded 11 distinct patterns (B to L) and a total of 28 subtypes. Representative SmaI digest patterns for the serotype V strains are shown in
Fig. 1. The most common PFGE pattern was
B (five subtypes), which was obtained with 60% of the isolates tested.
Indeed, 39 of the 64 isolates were highly related, having similarity
coefficients of >80% (data not shown). This pattern was genetically
related to the predominant U.S. serotype V GBS clone (pattern A) (Fig.
1). PFGE patterns C, D, F, G, H, K, and L were shared by four, three, four, two, two, four, and three isolates, respectively. PFGE patterns E, I, and J were each represented by a single isolate.
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.12.4526-4528.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Clonal Relationship between U.S. and French
Serotype V Group B Streptococcus Isolates
ABSTRACT
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80°C until further analysis was done.
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FIG. 1.
Representative major PFGE patterns of serotype V GBS
isolates. Lane 1, pattern A (predominant American clonal serotype V
GBS); lane 2, pattern B (39 isolates); lane 3, pattern C (4 isolates);
lane 4, pattern D (3 isolates); lane 5, pattern E (1 isolate); lane 6, pattern F (4 isolates); lane 7, pattern G (2 isolates); lane 8, pattern
H (2 isolates); lane 9, pattern I (1 isolate); lane 10, pattern J (1 isolate); lane 11, pattern K (4 isolates); lane 12, pattern L (3 isolates); lane 13, DNA size marker (Roche).
GBS have emerged as an important cause of morbidity and mortality among neonates, pregnant women, and other adults (9). In previous studies, serotypes Ia, Ib, II, and III were isolated from neonates with early-onset disease and from pregnant women with vaginal GBS colonization (4). Late-onset neonatal disease was due primarily to serotype III (25).
Serotype V GBS was first isolated in 1976 in the United States and was initially identified as NT1 (nontypeable type 1) (27); it was assigned type V status in 1985 (14). Serotype V appears to have emerged recently, because studies done before serotype V typing serum was available showed small percentages of nontypeable isolates (3). Serotype V seems to have been common since at least the mid-1980s in the United States (6) and also in Japan (23), Indonesia (26), The Gambia (22), Sweden (4), and France (1, 11, 16). Serotype V was the most common serotype recovered from nonpregnant adults with invasive GBS disease and the second and third most common serotype recovered from pregnant women and neonates with early-onset disease (6). French reports indicate that serotype V accounted for about 15% of isolates recovered from neonates (1, 16). Similar proportions (14 and 12%) of serotype V strains causing invasive infections were observed in neonates from Atlanta (6) and Maryland (13).
Molecular methods have been useful for discriminating among isolates of the same serotype (5, 7, 12). PFGE is a powerful technique for studying chromosomal relatedness among bacterial isolates. Blumberg et al. found that serotype V isolates recovered from patients in the Atlanta area were highly related (6). In our PFGE study, we obtained 11 patterns for the 64 serotype V GBS isolates, with one predominant. Genetic diversity among serotype V isolates had already been reported by Elliott et al. (8). However, these investigators have shown that 56% of 45 serotype V GBS isolates collected from 1986 to 1996 in the United States were genetically related (8). By PFGE, we found that this American clonal type, also found in Argentina (8), was indistinguishable from our predominant strain. Our study provides compelling evidence that the predominant isolates present in the United States and France are clonally related. Thus, the isolates in these two geographic locations are genetically diverse and have similar clonal structures.
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ACKNOWLEDGMENTS |
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We thank J. A. Elliott for providing the predominant American clonal serotype V GBS strain and D. Facklam for critical review of the manuscript.
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FOOTNOTES |
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* Corresponding author. Mailing address: Service de Microbiologie, Hôpital R. Debré, 48 Bd Sérurier, 75019 Paris, France. Phone: 33 (1) 40 03 23 40. Fax: 33 (1) 40 03 24 50. E-mail: edouard.bingen{at}rdb.ap-hop-paris.fr.
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Journal of Clinical Microbiology, December 2001, p. 4526-4528, Vol. 39, No. 12
0095-1137/01/$04.00+0 DOI: 10.1128/JCM.39.12.4526-4528.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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