![[Feedback]](/icons/banner/feedbackACT.gif)
![[For Subscribers]](/icons/banner/subscriberACT.gif)
![[Archive]](/icons/banner/archiveACT.gif)
![[Search]](/icons/banner/searchACT.gif)
![[Contents]](/icons/banner/tocACT.gif)
Previous Article | Next Article 
Journal of Clinical Microbiology, October 1999, p. 3411-3414, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Use of Multiplex PCR To Detect Classical and Newly Described
Pyrogenic Toxin Genes in Staphylococcal Isolates
Steven R.
Monday and
Gregory A.
Bohach*
Department of Microbiology, Molecular Biology
and Biochemistry, University of Idaho, Moscow, Idaho 83844
Received 21 April 1999/Returned for modification 8 June
1999/Accepted 14 July 1999
 |
ABSTRACT |
Staphylococcus aureus may contain one or more genes
that encode a variety of immunomodulatory pyrogenic toxins (PTs),
including the staphylococcal enterotoxins and toxic shock syndrome
toxin (TSST). The PTs interact with several cellular targets
to produce disease, such as food poisoning and toxic shock
syndrome. At present, nine serologically distinct
enterotoxins and one immunoreactive form of TSST have been identified
and characterized. As isolates of S. aureus are further
assessed, it is anticipated that this number will increase. To
facilitate screening, a multiplex PCR was designed to simultaneously
determine which of these 10 currently known PT genes an individual
S. aureus isolate possesses. We show here, using S. aureus isolates with characterized PT phenotypes, that this novel PCR technique reliably detects each of the known PTs in
a single reaction.
| |
TEXT |
Staphylococcus aureus is
a common pathogen that colonizes and produces disease in a variety of
hosts. The ability of this bacterium to successfully persist within
this range of hosts is largely due to the expression of a battery of
virulence factors which promote adhesion, acquisition of nutrients, and
evasion of host immunologic responses (18). Among these is
the pyrogenic exotoxin (PT) family, which is comprised of several
structurally and biologically related proteins expressed by
both S. aureus and Streptococcus
pyogenes (22).
PTs, which include toxic shock syndrome toxin (TSST) and the
staphylococcal enterotoxins (SEs), are secreted proteins that interact
with antigen-presenting cells and T lymphocytes to induce cellular
proliferation (12) and high-level cytokine expression (9). This activity does not involve the endocytic processing required for typical antigen presentation but instead occurs by concurrent association with major histocompatibility complex class II
molecules of the antigen-presenting cells and the V
domain of the
lymphocyte T-cell receptor (13). This interaction activates a much greater percentage of the host T-cell repertoire than that induced by antigens presented in the traditional manner
(15), explaining the massive cytokine expression and
subsequent immunomodulation brought about by these toxins.
Proteins which have the capacity to interact with the host
immunological system in this manner have been termed superantigens
(SAgs), and the PTs are prototypic examples of bacterial SAgs
(10).
The SEs and TSST are the causative agents of toxic shock syndrome
(6). Additionally, unlike the other members of the PT family, the SEs have the unique ability to induce staphylococcal food
poisoning, a common form of gastroenteritis (10).
Presently, nine major antigenic types of SEs have been reported
(SEA, SEB, SEC, SED, SEE, SEG, SEH, SEI, and SEJ), while only one
serotype of TSST (comprised of TSST-1 and TSSTovine) has
been described (1, 4, 5, 8, 19, 20, 24, 25). The SEC
serotype is heterogenous and contains several antigenic and
sequence molecular variants, designated SEC1, SEC2, SEC3,
SECbovine, and SECovine. These have been
classified on the basis of minor antigenic differences and the animal
host with which they are associated (17). Because of the
significance of these toxins for public health and food safety, an
efficient means for screening is needed. Also, since several of these
toxins have been discovered in very recent years, there is reason to
believe that as research on pathogenic S. aureus isolates
continues, additional SAgs will be described. Identification of novel
toxins will require an efficient means to screen isolates for
previously described staphylococcal PT genes. Toxigenic isolates that
do not harbor genes for currently recognized toxins are likely to
express novel SAgs. Therefore, we developed a multiplex PCR procedure
which will rapidly and simultaneously assess whether staphylococcal
isolates harbor sea, seb, sec,
sed, see, seg, seh, sei, and sej, encoding the SEs, and
tst, which encodes TSST.
Bacterial strains and DNA isolation.
Developing and testing of
the multiplex procedure was accomplished with DNA from the bacterial
strains listed in Table 1. Collectively,
these isolates contain all of the previously reported PT genes.
Staphylococcal genomic DNA was obtained from lysostaphin-treated cells
by a previously described process (7). The DNA was extracted with phenol and chloroform and was ethanol precipitated by standard methods (21). The DNA was recovered by centrifugation,
vacuum dried, resuspended in 200 µl of pyrogen-free H2O,
quantified spectrophotometrically at 260 and 280 nm, and diluted to a
final concentration of 10 ng/µl.
PCR primer design and amplification of bacterial DNA.
Nucleotide sequences for each of the PT genes were obtained from
GenBank by using their specific accession numbers (Table 2). The sequences were compared and
evaluated by using Genetics Computer Group (Madison, Wis.) computer
software to identify nucleotide sequences unique to each gene. With the
exception of the seb-sec primer set, which produces a 643-bp
amplification product common to both seb and sec,
all primer sets were designed to anneal to unique regions and generate
amplification products that would allow identification of each PT gene
based on the molecular weight of its PCR product (Table 2). To
discriminate whether the 643-bp seb-sec amplification
product is indicative of either seb or sec, a
separate 5' sec primer was designed that works in
combination with the 3' seb-sec primer to produce a 283-bp
amplification product unique to sec, including the bovine
variant (see below). Additionally, to ensure that toxin-negative
samples were interpreted correctly and that a sufficient quantity of
PCR template DNA was present, the samples were also tested by PCR with
a primer set that anneals to the S. aureus 16S rRNA gene
that generates a 228-bp amplicon during the amplification process
(Table 2).
The multiplex PCR was performed in a 50-µl volume with the Gibco BRL
Taq DNA polymerase system (Life Technologies, Inc.,
Rockville, Md.) containing the following: 1× Taq polymerase
buffer, 4 mM MgCl2, 300 nM concentrations of each of the
primers listed in Table 2, 400 µM concentrations of deoxynucleoside
triphosphates, 5 U of Taq polymerase, and 50 ng of
staphylococcal DNA. Bulk solutions containing the Taq
buffer, MgCl2, deoxynucleoside triphosphates, and multiplex
PCR primer mix at the appropriate concentrations were prepared for the
desired number of reactions. Aliquots of the staphylococcal template
DNA (50 ng in 5 µl) from each test strain were individually placed
into 500-µl thin-walled PCR tubes. Afterwards, 40 µl of the bulk
solution was added to each tube containing the template DNA and covered
with 50 µl of mineral oil. These tubes were subsequently incubated
for 10 min (95°C), during which (after the initial 3 min) 5 U of
Taq polymerase (in 5 µl [total volume] of 1×
Taq buffer) was added to each reaction. Following this
"hot-start" procedure, DNA was amplified in an Amplitron II
thermocycler (Barnstead Thermolyne Co., Dubuque, Iowa) by 15 cycles of
95°C for 1 min, 68°C for 45 s, and 72°C for 1 min and 16 cycles of 95°C for 1 min, 64°C for 45 s, and 72°C for 1 min.
The reaction was terminated with a 10-min incubation at 72°C.
PCR products were resolved by electrophoresis in 1.5% agarose (0.5×
Tris boric acid, EDTA) gels at 100 V (constant voltage) and visualized
on a transilluminator with a charged coupled device camera and the
Molecular Analyst software (Bio-Rad, Hercules, Calif.). Product sizes
were determined by using the 1 Kb Plus DNA molecular weight ladder
(Life Technologies, Inc.).
Genetic analysis of clinical strains using the multiplex PCR
procedure.
Analyses using DNA obtained from the staphylococcal
isolates demonstrated that all primer pairs produced amplification
products consistent with their predicted sizes. For example,
amplification of DNA from FRI913, known to produce SEA, SEC, SEE and
TSST-1 (3), generated bands indicative of sea
(520 bp), sec (643 and 283 bp), see (170 bp), and
tst (559 bp) (Fig. 1).
Similarly, amplification of DNA obtained from S. aureus MN
HOCH (7), FRI472 (16), FRI326 (16),
FRI572 (19), FRI569 (24), and FRI445
(19) produced PCR products consistent with the toxin genes
that had previously been reported for these strains (Table 1 and Fig. 1). As expected, while amplification of DNA obtained from the nontoxigenic strain RN4220 consistently produced the 16S rRNA gene
amplicon (results not shown), it failed to produce any SE-related PCR
product when subjected to the multiplex PCR analysis. Since a
single isolate may contain multiple toxin genes, we assessed the
number of genes which could be simultaneously detected with this
process under these particular reaction conditions. Using DNA pooled
from several isolates (FRI913, FRI472, and FRI569), we demonstrated
that it was possible to simultaneously generate PCR products
representative of all 10 PT genes, as well as the 16S rRNA gene, in a
single reaction (results not shown).
View larger version (51K):
[in this window]
[in a new window]
|
FIG. 1.
Agarose gel electrophoresis of the multiplex PCR
amplification products from analysis of bacterial test strains. Lanes
contain amplification products of DNA isolated from strains designated
at top. Product identification was facilitated by direct comparison to
the 1 Kb Plus (Gibco BRL) molecular weight ladder (far left lane) and a
PT gene amplification product ladder prepared by pooling reaction
products generated in individual reactions (far right lane).
|
|
By design, amplification of DNA obtained from strains producing SEC
were expected to result in the production of the 643- and 283-bp
amplification products indicative of sec, like that observed
for FRI913 (Fig. 1). However, strains known to express SECbovine did not generate both amplicons when tested by
this procedure. Amplification of DNA obtained from the
SECbovine+ strain 3169 (Fig. 1) (17)
resulted in the production of the 283-bp amplicon but did not produce
the 643-bp seb-sec PCR product typically observed in
analyses of SEB- or SEC-expressing isolates (MN HOCH and FRI913,
respectively) (Fig. 1). This finding, which was also observed when DNA
isolated from another known SECbovine-producing S. aureus isolate was tested (results not shown), suggests that the
gene encoding SECbovine has unique attributes not shared by other sec genes (see below). The 5' seb-sec
primer was designed to anneal to these genes at a conserved sequence
within the region encoding the signal peptides of SEB and SEC. Since
amplification of DNA derived from SECbovine producers does
not result in the production of the 643-bp amplicon, it is likely that
the region of the gene encoding the SECbovine signal
peptide is comprised of a nucleotide sequence dissimilar to that of the
other sec variants. At present, the nucleotide sequence
within this portion of the SECbovine gene is not available
in GenBank and, therefore, cannot be directly compared to analogous
regions of the other sec variants.
The multiplex PCR process described in this report reliably detects the
genes for all staphylococcal PTs reported as of April 1999. Moreover,
this technique was shown to be able to detect at least four different
SE genes (producing five separate PCR products) in a single bacterial
isolate. The fact that it can simultaneously detect all 10 genes in a
pooled sample of DNA ensures that this procedure can both confirm the
presence of PT genes previously associated with a particular strain and
detect other currently known toxin genes within the isolate. For
instance, in the present study S. aureus FRI472, previously
described as a SED producer (1), consistently generated a
384-bp PCR product, confirming the presence of the sed gene
within its genome. However, amplicons with molecular sizes of 143, 327, and 465 bp were also produced, indicating that FRI472 contains
genes encoding SEJ, SEG, and SEI, respectively, as well (Fig. 1). This
observation is consistent with the recent report by Zhang et al.
(25), who determined that the SEJ determinant is present on
the same plasmid as the SED determinant. Similarly, we also showed that
other isolates, such as S. aureus FRI572 and FRI445, carry
toxin genes not previously associated with these isolates (Fig. 1).
This work has produced a system that expands the capabilities of the
multiplex PCR procedures previously developed by several other
investigators (2, 11, 14, 23). Most notably, the system
described in this report reliably, rapidly, and simultaneously detects
each of the 10 currently described staphylococcal toxin determinants,
including the most recently described seg (19), sei (19), and sej (25)
genes. Additionally, in a single reaction, the process generates
amplicons that allow easy discrimination of the determinants an isolate
carries, regardless of the number of PT genes carried by the isolate.
These features allow this procedure to be applied in the clinical
setting for epidemiological studies or to guide therapeutic strategies.
This efficient method of screening isolates for PT genes could also
facilitate the identification of additional genes encoding novel,
yet-undescribed toxins.
| |
ACKNOWLEDGMENTS |
This work was supported by the funds provided by Public Health
Service grant AI28401, the United Dairymen of Idaho, and the Idaho
Agriculture Experiment Station (G.A.B.).
We thank Claudia Deobald for her assistance in the development of this
assay. We also thank Patrick Schlievert (at the University of
Minnesota), Amy Wong (at the Food Research Institute at the University
of Wisconsin), and Merlin Bergdoll (also at the Food Research
Institute) for kindly providing many of the strains used in this work.
| |
FOOTNOTES |
*
Corresponding author. Mailing address: Department of
Microbiology, Molecular Biology and Biochemistry, University of Idaho, Moscow, ID 83844. Phone: (208) 885-6666. Fax: (208) 885-6518. E-mail:
gbohach{at}uidaho.edu.
| |
REFERENCES |
| 1.
|
Bayles, K. W., and J. J. Iandolo.
1989.
Genetic and molecular analyses of the gene encoding staphylococcal enterotoxin D.
J. Bacteriol.
171:4799-4806[Abstract/Free Full Text].
|
| 2.
|
Becker, K.,
R. Roth, and G. Peters.
1998.
Rapid and specific detection of toxinogenic Staphylococcus aureus: use of two multiplex PCR enzyme immunoassays for amplification and hybridization of staphylococcal enterotoxin genes, exfoliative toxin genes, and toxic shock syndrome toxin 1 gene.
J. Clin. Microbiol.
36:2548-2553[Abstract/Free Full Text].
|
| 3.
| Bergdoll, M. S. 1988. Personal communication.
|
| 4.
|
Betley, M. J., and J. J. Mekalanos.
1988.
Nucleotide sequence of the type A staphylococcal enterotoxin gene.
J. Bacteriol.
170:34-41[Abstract/Free Full Text].
|
| 5.
|
Bohach, G. A.
1997.
Staphylococcal enterotoxins B and C. Structural requirements for superantigenic and enterotoxigenic activities.
Prep. Biochem. Biotechnol.
27:79-110[Medline].
|
| 6.
|
Bohach, G. A.,
D. J. Fast,
R. D. Nelson, and P. M. Schlievert.
1990.
Staphylococcal and streptococcal pyrogenic toxins involved in toxic shock syndrome and related illnesses.
Crit. Rev. Microbiol.
17:251-272[Medline].
|
| 7.
|
Bohach, G. A.,
B. N. Kreiswirth,
R. P. Novick, and P. M. Schlievert.
1989.
Analysis of toxic shock syndrome isolates producing staphylococcal enterotoxins B and C1 with use of southern hybridization and immunologic assays.
Rev. Infect. Dis.
11(Suppl.1):S75-S81.
|
| 8.
|
Couch, J. L.,
M. T. Soltis, and M. J. Betley.
1988.
Cloning and nucleotide sequence of the type E staphylococcal enterotoxin gene.
J. Bacteriol.
170:2954-2960[Abstract/Free Full Text].
|
| 9.
|
Fast, D. J.,
P. M. Schlievert, and R. D. Nelson.
1988.
Nonpurulent response to toxic shock syndrome toxin 1-producing Staphylococcus aureus. Relationship to toxin stimulated production of tumor necrosis factor.
J. Immunol.
140:949-953[Abstract].
|
| 10.
|
Jablonski, L. M., and G. A. Bohach.
1997.
Staphylococcus aureus, p. 353-375.
In
M. P. Doyle, L. P. Beuchat, and T. J. Montville (ed.), Food microbiology: fundamentals and frontiers. ASM Press, Washington, D.C.
|
| 11.
|
Johnson, W. M.,
S. D. Tyler,
E. P. Ewan,
F. E. Ashton,
D. R. Pollard, and K. R. Rozee.
1991.
Detection of genes for enterotoxins, exfoliative toxins, and toxic shock syndrome toxin 1 in Staphylococcus aureus by the polymerase chain reaction.
J. Clin. Microbiol.
29:426-430[Abstract/Free Full Text].
|
| 12.
|
Kotb, M.
1995.
Bacterial pyrogenic exotoxins as superantigens.
Clin. Microbiol. Rev.
8:411-426[Abstract].
|
| 13.
|
Lavoie, P. M.,
R. P. Sekaly,
J. Thibodeau, and F. Denis.
1997.
Interaction of superantigens with MHC class II molecules, p. 61-83.
In
D. Y. Leung, B. T. Huber, and P. Schlievert (ed.), Superantigens: molecular biology, immunology, and relevance to human disease. Marcel Dekker, Inc., New York, N.Y.
|
| 14.
|
Lee, S. U.,
M. Quesnell,
L. K. Fox,
J. W. Yoon,
Y. H. Park,
W. C. Davis,
D. Falk,
C. F. Deobald, and G. A. Bohach.
1998.
Characterization of staphylococcal bovine mastitis isolates using polymerase chain reaction.
J. Food Prot.
61:1384-1386.
[Medline] |
| 15.
|
Leung, D. Y.,
B. T. Huber, and P. Schlievert.
1997.
Historical perspectives of superantigens and their biological activities, p. 1-13.
In
D. Y. Leung, B. T. Huber, and P. Schlievert (ed.), Superantigens: molecular biology, immunology, and relevance to human disease. Marcel Dekker, Inc., New York, N.Y.
|
| 16.
|
Louch, H. A.,
M. L. Eck, and K. J. Miller.
1997.
Osmoadaptation by Staphylococcus aureus; analysis of several strains linked to food poisoning outbreaks.
J. Food Prot.
60:139-143.
|
| 17.
|
Marr, J. C.,
J. D. Lyon,
J. R. Roberson,
M. Lupher,
W. C. Davis, and G. A. Bohach.
1993.
Characterization of novel type C staphylococcal enterotoxins: biological and evolutionary implications.
Infect. Immun.
61:4254-4262[Abstract/Free Full Text].
|
| 18.
|
Monday, S. R., and G. A. Bohach.
1999.
Properties of Staphylococcus aureus enterotoxins and toxic shock syndrome toxin-1, p. 589-610.
In
J. E. Alouf, and J. H. Freer (ed.), The comprehensive sourcebook of bacterial protein toxins, 2nd ed. Academic Press, London, England.
|
| 19.
|
Munson, S. H.,
M. T. Tremaine,
M. J. Betley, and R. A. Welch.
1998.
Identification and characterization of staphylococcal enterotoxin types G and I from Staphylococcus aureus.
Infect. Immun.
66:3337-3348[Abstract/Free Full Text].
|
| 20.
|
Murray, D. L.,
G. S. Prasad,
C. A. Earhart,
B. A. B. Leonard,
B. N. Kreiswirth,
R. P. Novick,
D. H. Ohlendorf, and P. M. Schlievert.
1994.
Immunobiologic and biochemical properties of mutants of toxic shock syndrome toxin-1.
J. Immunol.
152:87-95[Abstract].
|
| 21.
|
Sambrook, J.,
E. F. Fritsch, and T. Maniatis.
1989.
Molecular cloning: a laboratory manual.
Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
|
| 22.
|
Schlievert, P. M.,
G. A. Bohach,
D. H. Ohlendorf,
C. V. Stauffacher,
D. Y. Leung,
D. L. Murray,
G. S. Prasad,
C. A. Earhart,
L. M. Jablonski,
M. L. Hoffmann, and Y. I. Chi.
1995.
Molecular structure of Staphylococcus and Streptococcus superantigens.
J. Clin. Immunol.
15:4S-10S[Medline]. (Erratum, 16:126, 1996.)
|
| 23.
|
Schmitz, F. J.,
M. Steiert,
B. Hofmann,
J. Verhoef,
U. Hadding,
H. P. Heinz, and K. Kohrer.
1998.
Development of a multiplex-PCR for direct detection of the genes for enterotoxin B and C, and toxic shock syndrome toxin-1 in Staphylococcus aureus.
J. Med. Microbiol.
47:335-340[Abstract].
|
| 24.
|
Su, Y. C., and A. C. Wong.
1995.
Identification and purification of a new staphylococcal enterotoxin, H.
Appl. Environ. Microbiol.
61:1438-1443[Abstract].
|
| 25.
|
Zhang, S.,
J. J. Iandolo, and G. C. Stewart.
1998.
The enterotoxin D plasmid of Staphylococcus aureus encodes a second enterotoxin determinant (sej).
FEMS Microbiol. Lett.
168:227-233[Medline].
|
Journal of Clinical Microbiology, October 1999, p. 3411-3414, Vol. 37, No. 10
0095-1137/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Parsonnet, J., Goering, R. V., Hansmann, M. A., Jones, M. B., Ohtagaki, K., Davis, C. C., Totsuka, K.
(2008). Prevalence of Toxic Shock Syndrome Toxin 1 (TSST-1)-Producing Strains of Staphylococcus aureus and Antibody to TSST-1 among Healthy Japanese Women. J. Clin. Microbiol.
46: 2731-2738
[Abstract]
[Full Text]
-
Spence, R. P., Wright, V., Ala-Aldeen, D. A. A., Turner, D. P., Wooldridge, K. G., James, R.
(2008). Validation of Virulence and Epidemiology DNA Microarray for Identification and Characterization of Staphylococcus aureus Isolates. J. Clin. Microbiol.
46: 1620-1627
[Abstract]
[Full Text]
-
Sauer, P., Sila, J., Stosova, T., Vecerova, R., Hejnar, P., Vagnerova, I., Kolar, M., Raclavsky, V., Petrzelova, J., Loveckova, Y., Koukalova, D.
(2008). Prevalence of genes encoding extracellular virulence factors among meticillin-resistant Staphylococcus aureus isolates from the University Hospital, Olomouc, Czech Republic. J Med Microbiol
57: 403-410
[Abstract]
[Full Text]
-
Collery, M. M., Smyth, D. S., Twohig, J. M., Shore, A. C., Coleman, D. C., Smyth, C. J.
(2008). Molecular typing of nasal carriage isolates of Staphylococcus aureus from an Irish university student population based on toxin gene PCR, agr locus types and multiple locus, variable number tandem repeat analysis. J Med Microbiol
57: 348-358
[Abstract]
[Full Text]
-
Strommenger, B., Braulke, C., Pasemann, B., Schmidt, C., Witte, W.
(2008). Multiplex PCR for Rapid Detection of Staphylococcus aureus Isolates Suspected to Represent Community-Acquired Strains. J. Clin. Microbiol.
46: 582-587
[Abstract]
[Full Text]
-
Bonness, S., Szekat, C., Novak, N., Bierbaum, G.
(2008). Pulsed-Field Gel Electrophoresis of Staphylococcus aureus Isolates from Atopic Patients Revealing Presence of Similar Strains in Isolates from Children and Their Parents. J. Clin. Microbiol.
46: 456-461
[Abstract]
[Full Text]
-
Sasaki, T., Kikuchi, K., Tanaka, Y., Takahashi, N., Kamata, S., Hiramatsu, K.
(2007). Methicillin-Resistant Staphylococcus pseudintermedius in a Veterinary Teaching Hospital. J. Clin. Microbiol.
45: 1118-1125
[Abstract]
[Full Text]
-
Collery, M. M., Smyth, C. J.
(2007). Rapid differentiation of Staphylococcus aureus isolates harbouring egc loci with pseudogenes {psi}ent1 and {psi}ent2 and the selu or seluv gene using PCR-RFLP. J Med Microbiol
56: 208-216
[Abstract]
[Full Text]
-
Kwon, N. H., Park, K. T., Moon, J. S., Jung, W. K., Kim, S. H., Kim, J. M., Hong, S. K., Koo, H. C., Joo, Y. S., Park, Y. H.
(2005). Staphylococcal cassette chromosome mec (SCCmec) characterization and molecular analysis for methicillin-resistant Staphylococcus aureus and novel SCCmec subtype IVg isolated from bovine milk in Korea. J Antimicrob Chemother
56: 624-632
[Abstract]
[Full Text]
-
Dakic, I., Vukovic, D., Stepanovic, S., Hauschild, T., Jezek, P., Petras, P., Morrison, D.
(2005). Survey of Genes Encoding Staphylococcal Enterotoxins, Toxic Shock Syndrome Toxin 1, and Exfoliative Toxins in Members of the Staphylococcus sciuri Group. J. Clin. Microbiol.
43: 4875-4876
[Abstract]
[Full Text]
-
Holmes, A., Ganner, M., McGuane, S., Pitt, T. L., Cookson, B. D., Kearns, A. M.
(2005). Staphylococcus aureus Isolates Carrying Panton-Valentine Leucocidin Genes in England and Wales: Frequency, Characterization, and Association with Clinical Disease. J. Clin. Microbiol.
43: 2384-2390
[Abstract]
[Full Text]
-
Smyth, D. S, Hartigan, P. J, Meaney, W. J, Fitzgerald, J R., Deobald, C. F, Bohach, G. A, Smyth, C. J
(2005). Superantigen genes encoded by the egc cluster and SaPIbov are predominant among Staphylococcus aureus isolates from cows, goats, sheep, rabbits and poultry. J Med Microbiol
54: 401-411
[Abstract]
[Full Text]
-
Cassat, J. E., Dunman, P. M., McAleese, F., Murphy, E., Projan, S. J., Smeltzer, M. S.
(2005). Comparative Genomics of Staphylococcus aureus Musculoskeletal Isolates. J. Bacteriol.
187: 576-592
[Abstract]
[Full Text]
-
Lovseth, A., Loncarevic, S., Berdal, K. G.
(2004). Modified Multiplex PCR Method for Detection of Pyrogenic Exotoxin Genes in Staphylococcal Isolates. J. Clin. Microbiol.
42: 3869-3872
[Abstract]
[Full Text]
-
Velazquez-Meza, M. E., Aires de Sousa, M., Echaniz-Aviles, G., Solorzano-Santos, F., Miranda-Novales, G., Silva-Sanchez, J., de Lencastre, H.
(2004). Surveillance of Methicillin-Resistant Staphylococcus aureus in a Pediatric Hospital in Mexico City during a 7-Year Period (1997 to 2003): Clonal Evolution and Impact of Infection Control. J. Clin. Microbiol.
42: 3877-3880
[Abstract]
[Full Text]
-
El-Zimaity, D., Kearns, A. M., Dawson, S. J., Price, S., Harrison, G. A. J.
(2004). Survey, characterization and susceptibility to fusidic acid of Staphylococcus aureus in the Carmarthen area. J Antimicrob Chemother
54: 441-446
[Abstract]
[Full Text]
-
Carneiro, L. A.M., Queiroz, M. L.P., Merquior, V. L.C.
(2004). Antimicrobial-resistance and enterotoxin-encoding genes among staphylococci isolated from expressed human breast milk. J Med Microbiol
53: 761-768
[Abstract]
[Full Text]
-
Holtfreter, S., Bauer, K., Thomas, D., Feig, C., Lorenz, V., Roschack, K., Friebe, E., Selleng, K., Lovenich, S., Greve, T., Greinacher, A., Panzig, B., Engelmann, S., Lina, G., Broker, B. M.
(2004). egc-Encoded Superantigens from Staphylococcus aureus Are Neutralized by Human Sera Much Less Efficiently than Are Classical Staphylococcal Enterotoxins or Toxic Shock Syndrome Toxin. Infect. Immun.
72: 4061-4071
[Abstract]
[Full Text]
-
Sergeev, N., Volokhov, D., Chizhikov, V., Rasooly, A.
(2004). Simultaneous Analysis of Multiple Staphylococcal Enterotoxin Genes by an Oligonucleotide Microarray Assay. J. Clin. Microbiol.
42: 2134-2143
[Abstract]
[Full Text]
-
Aucken, H. M., Ganner, M., Murchan, S., Cookson, B. D., Johnson, A. P.
(2002). A new UK strain of epidemic methicillin-resistant Staphylococcus aureus (EMRSA-17) resistant to multiple antibiotics. J Antimicrob Chemother
50: 171-175
[Abstract]
[Full Text]
-
Oliveira, D. C., Lencastre, H. d.
(2002). Multiplex PCR Strategy for Rapid Identification of Structural Types and Variants of the mec Element in Methicillin-Resistant Staphylococcus aureus. Antimicrob. Agents Chemother.
46: 2155-2161
[Abstract]
[Full Text]
-
Omoe, K., Ishikawa, M., Shimoda, Y., Hu, D.-L., Ueda, S., Shinagawa, K.
(2002). Detection of seg, seh, and sei genes in Staphylococcus aureus Isolates and Determination of the Enterotoxin Productivities of S. aureus Isolates Harboring seg, seh, or sei Genes. J. Clin. Microbiol.
40: 857-862
[Abstract]
[Full Text]
-
Akineden, O., Annemuller, C., Hassan, A. A., Lammler, C., Wolter, W., Zschock, M.
(2001). Toxin Genes and Other Characteristics of Staphylococcus aureus Isolates from Milk of Cows with Mastitis. CVI
8: 959-964
[Abstract]
[Full Text]
-
Moore, P. C. L., Lindsay, J. A.
(2001). Genetic Variation among Hospital Isolates of Methicillin-Sensitive Staphylococcus aureus: Evidence for Horizontal Transfer of Virulence Genes. J. Clin. Microbiol.
39: 2760-2767
[Abstract]
[Full Text]
-
Fitzgerald, J. R., Monday, S. R., Foster, T. J., Bohach, G. A., Hartigan, P. J., Meaney, W. J., Smyth, C. J.
(2001). Characterization of a Putative Pathogenicity Island from Bovine Staphylococcus aureus Encoding Multiple Superantigens. J. Bacteriol.
183: 63-70
[Abstract]
[Full Text]
-
Sharma, N. K., Rees, C. E. D., Dodd, C. E. R.
(2000). Development of a Single-Reaction Multiplex PCR Toxin Typing Assay for Staphylococcus aureus Strains. Appl. Environ. Microbiol.
66: 1347-1353
[Abstract]
[Full Text]
Top
Abstract
Text
