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Journal of Clinical Microbiology, November 1999, p. 3759-3760, Vol. 37, No. 11
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
A Serotype VIII Strain among Colonizing Group B Streptococcal
Isolates in Boston, Massachusetts
Leanne J.
Paoletti,1
Jessica
Bradford,2 and
Lawrence C.
Paoletti2,3,*
Clinical Microbiology
Department1 and Channing Laboratory,
Department of Medicine,2 Brigham and
Women's Hospital, and Harvard Medical
School,3 Boston, Massachusetts
Received 24 June 1999/Returned for modification 22 July
1999/Accepted 16 August 1999
 |
ABSTRACT |
Maternal colonization with group B Streptococcus (GBS)
is a risk factor for neonatal GBS disease. Whereas serotypes Ia, Ib, II, III, and V are prevalent in the United States, types VI and VIII
predominate in Japan. Recently, a serotype VIII strain was detected
among 114 clinical GBS isolates from a Boston, Mass., hospital.
| |
TEXT |
Group B Streptococcus
(GBS) is a prominent pathogen responsible for sepsis, pneumonia, and
meningitis among newborns; urinary tract infection, endometritis, and
chorioamnionitis among peripartum women; and primary bacteremia among
the elderly (12). Although GBS is a component of the normal
human gastrointestinal flora, vaginal and/or rectal colonization with
GBS during pregnancy is a risk factor for neonatal GBS disease. Of the
nine currently identified GBS serotypes, five (Ia, Ib, II, III, and V)
are prevalent in the United States, with type V as the most recently
emergent serotype (2, 11). GBS types VI and VIII predominate
among pregnant Japanese women (4), and only a single isolate
of type VIII has thus far been reported in the United States
(2). GBS types IV and VII are encountered only rarely
(2, 3). Distribution studies reveal population shifts of
known GBS serotypes and the emergence of new types; these changes have
implications for the formulation of a multivalent vaccine. In this
study, we examined the present distribution of GBS serotypes in Boston,
Mass., where the first serotype distribution study was performed over
35 years ago (1).
One hundred nine clinical samples obtained between January and March
1999 from the Brigham and Women's Hospital Clinical Microbiology Laboratory were identified as belonging to Lancefield's group B by a
latex agglutination test (Streptex; Murex Diagnostics, Inc., Norcross,
Ga.). GBS isolates were individually scored on blood agar plates and
incubated overnight at 37°C. Included on each plate as positive
controls were GBS strains of type Ia (O90), type Ib (H36B), type II
(18RS21), type III (M781), and type V (CJB111). Each isolate was
sequentially transferred from the blood agar plate onto five
nitrocellulose membranes (Millipore Corporation, Bedford, Mass.) by
gently placing the membranes over the colonies for 5 to 10 s. The
membranes were then placed (sample side up) into an empty petri plate,
and the samples were heat fixed by incubation at 60°C for 1 h.
Membranes were blocked with 1% skim milk and rinsed thrice with 40 mM
phosphate buffer containing 0.05% Tween 20 (PB/T). Rabbit serum
specific to each GBS capsular polysaccharide (CPS)-tetanus toxoid
conjugate vaccine, i.e., serotype Ia, Ib, II, III, V, VI, and VIII
conjugates (8-10, 13, 14), served as the primary antibody
and was used at a dilution at which cross-reactions with heterologous
serotypes were minimal to none. The membranes were incubated with the
primary antibody for 1 h at room temperature with gentle agitation
and then washed thrice with PB/T. Goat anti-rabbit immunoglobulin
G-alkaline phosphatase conjugate (Southern Biotechnology
Associates, Birmingham, Ala.), diluted 1:3,000, was used as the
secondary antibody; after its addition, the membranes were incubated
for 1 h with gentle agitation at room temperature. Membranes were
rinsed thrice with PB/T and developed by the addition of
5-bromo-4-chloro-3-indolylphosphate toluidinium-nitroblue tetrazolium
phosphatase substrate (Kirkegaard and Perry Laboratories, Gaithersburg,
Md.). The reaction was stopped by rinsing the membranes with water.
Ninety-four percent (103 of 109) of the GBS samples were cultured from
females; and two isolates were obtained from newborns. Most (90.7%) of
the isolates originated from the genitourinary tract of women. A total
of 114 GBS isolates were obtained from the 109 samples, with dual
serotypes in five samples (4.4%). GBS types Ia, III, and V accounted
for 80% of the isolates (Fig. 1), and
types Ib and II accounted for 15%. Surprisingly, one isolate was
identified as type VIII
a serotype rare outside Japan (2). Only four isolates (3.5%) were nontypeable.
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FIG. 1.
Comparison of group B streptococcal serotype
distribution in Boston among 114 clinical isolates obtained in 1999 (black bars) and among 149 isolates obtained in 1962 to 1963 (white
bars) (1).
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|
Samples that reacted strongly with antibody to more than one serotype
were again plated, and 10 to 15 individual colonies were grown on a
blood agar plate, transferred to nitrocellulose membranes, and probed
with specific antiserum to the serotypes present in the first screen.
An interesting finding was that type V was isolated from each
dual-serotype sample (Ia-V, n = 1; Ib-V, n = 2; III-V, n = 2). Moreover, type V was found in
three of the four samples from males (type III was isolated from the
fourth sample). Of the two isolates from babies, one was type Ia and the other was type III.
In the past, serotyping of GBS has required a tedious process of
antigen extraction and precipitation reactions with adsorbed whole-cell
antisera (5). Adsorption of whole-cell antisera was
necessary to remove cross-reactivity with other GBS surface components,
such as the group B carbohydrate and protein antigens. The availability
of high-titered CPS-specific antisera to conjugate vaccines has
resulted in a more direct and rapid method of serotyping while
eliminating the need for extensive adsorptions to remove antibody to
non-CPS surface antigens; the minor CPS cross-reactivity in some of the
high-titered antisera has been reduced by adsorption with the
interfering GBS serotype. High-titered CPS-specific sera also have
allowed the detection of more than one serotype in a sample. In this
study, each of five samples contained two serotypes, and type V was
involved in each of these cases.
A study by Eickhoff and colleagues in Boston in 1964 revealed that
collectively types Ia and III accounted for 68% of all cases of GBS
infection (1), whereas in our study, serotypes Ia and III
together accounted for 49% of the total (Fig. 1). Serotype V had not
emerged in the early 1960s but accounted for 32% of the total isolates
in our study (Fig. 1). Indeed, the overall distribution of GBS
serotypes in our study was similar to those in current, and more
extensively performed, epidemiological studies (2, 6).
However, of the combined 614 invasive pediatric and adult isolates
collected in those studies, only one was serotype VIII. The occurrence
of GBS type VIII among our relatively small number of samples may
signal future changes in serotype distribution in the United States. A
multivalent GBS vaccine for use in the United States would currently
include types Ia, Ib, II, III, and V (7); this formulation
would provide coverage against 96% of the isolates in this study.
| |
ACKNOWLEDGMENTS |
We thank Andrew B. Onderdonk and Dennis L. Kasper for advice and
critical review of the manuscript and Julie McCoy and Jaylyn Olivo for
editorial assistance.
This work was supported by an award from the William F. Milton Fund to
L.C.P.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Channing
Laboratory, 181 Longwood Ave., Boston, MA 02115. Phone: (617) 525-2678. Fax: (617) 731-1541. E-mail:
lpaoletti{at}channing.harvard.edu.
| |
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Journal of Clinical Microbiology, November 1999, p. 3759-3760, Vol. 37, No. 11
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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