Association of toxic shock toxin-1 determinant with a heterologous insertion at multiple loci in the Staphylococcus aureus chromosome

Chu, May C.; Kreiswirth, Barry N.; Pattee, P A; Novick, Richard P.; Melish, Marian E.; James, J. F.

doi:10.1128/iai.56.10.2702-2708.1988

Cited by 36 publications

(8 citation statements)

References 40 publications

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“…5, tracks L, B, F, G. J, K. N, R. S, V, W, X) as did the Hla" TSST-1' Harnsburg strain (data not shown). The large HindUl fragment hybridizing to the tst probe is associated with strains that are tryptophan-requinng and have the (sf element located close to the trp locus (Chu et al, 1988;Kreiswirth ef al., 1989).…”

Section: Hybridization Analysis Ot Clinical Isolatesmentioning

confidence: 99%

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Cryptic α‐toxin gene in toxic shock syndrome and septicaemia strains of Staphylococcus aureus

O’Reilly

Kreiswirth²,

Foster

1990

Molecular Microbiology

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The majority of clinical isolates of Staphylococcus aureus that produce toxic shock syndrome toxin-1 (TSST-1) fail to express alpha-toxin, despite having a copy of the hla gene in the chromosome. The hla gene was cloned from an Hla- TSST-1+ strain, Todd 555, which had been isolated from a case of toxic shock syndrome in the USA. Of the 630 bases of the Todd 555 gene sequenced, 46 differed from the hla gene sequence of strain Wood 46. The defect in alpha-toxin expression was shown to be due to a nonsense mutation which converted a CAG glutamine codon in the equivalent position in the functional Wood 46 sequence to a TAG stop codon. The same mutation was present in the hla gene cloned from a human septicaemia strain (V37) isolated in Dublin. The nonsense mutation of Todd 555 was suppressed by the supE44 mutation in Escherichia coli resulting in haemolytic activity in cell lysates. Hybrid hla genes were formed by splicing fragments of hla from Todd 555 and Wood 46. Expression of one such chimaeric hla gene in S. aureus demonstrated that the Todd 555 hla gene has a functional agr-regulated promoter. The silent hla gene may be a cryptic gene in S. aureus.

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Section: Hybridization Analysis Ot Clinical Isolatesmentioning

confidence: 99%

“…5, track H) did not co-migrate with the tst fragment of other TSST-1-producing isolates. It has previously been shown that restriction fragment-length polymorphisms (RFLPs) associated with fs( reflect different chromosomal locations (Chu et al. 1988; Kreiswirth ef a/., 1989).…”

Section: Hybridization Analysis Ot Clinical Isolatesmentioning

confidence: 99%

Cryptic α‐toxin gene in toxic shock syndrome and septicaemia strains of Staphylococcus aureus

O’Reilly

Kreiswirth²,

Foster

1990

Molecular Microbiology

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“…(1983) and later shown to be a chromosomal gene embedded in an accessory genetic element that contains conserved tst flanking sequences and is entirely absent from TSST‐1‐negative strains (Kreiswirth et al ., 1983; 1989). Tst is located near tyrB in strain RN4282, the source strain for the original clone, which belongs to agr peptide group I (Ji et al ., 1997), and within the trp locus in most other menstrual TSS isolates (Chu et al ., 1988), which belong to agr peptide group III (Ji et al ., 1997). It therefore seemed likely, at the time the present studies were initiated, that the tst element had inserted into two different sites and was therefore a transposon.…”

Section: Introductionmentioning

confidence: 99%

The gene for toxic shock toxin is carried by a family of mobile pathogenicity islands in Staphylococcus aureus

Lindsay

Ruzin

Ross

et al. 1998

Molecular Microbiology

398

356

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SummaryTst, the gene for toxic shock syndrome toxin-1 (TSST-1), is part of a 15.2 kb genetic element in Staphylococcus aureus that is absent in TSST-1-negative strains. The prototype, in RN4282, is flanked by a 17 nucleotide direct repeat and contains genes for a second possible superantigen toxin, a Dichelobacter nodosus VapE homologue and a putative integrase. It is readily transferred to a recA ¹ recipient, and it always inserts into a unique chromosomal copy of the 17 nucleotide sequence in the same orientation. It is excised and circularized by staphylococcal phages 13 and 80␣ and replicates during the growth of the latter, which transduces it at very high frequency. Because of its site and orientation specificity and because it lacks other identifiable phage-like genes, we consider it to be a pathogenicity island (PI) rather than a transposon or a defective phage. The tst element in RN4282, near tyrB, is designated SaPI1. That in RN3984 in the trp region is only partially homologous to SaPI1 and is excised by phage 80 but not by 80␣. It is designated SaPI2. These PIs are the first in any Gram-positive species and the first for which mobility has been demonstrated. Their mobility may be responsible for the spread of TSST-1 production among S. aureus strains.

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“…F(ab')2 fragments of IgG antibodies do not bind to protein A (26) and have been used successfully in ELISA procedures (16). Moreover, the generation of F(ab')2 fragments is simple and rapid, since pepsin digestion of whole IgG and purification of F(ab')2 can be accomplished within 24 h. In addition to the use of F(ab')2, other methods have been used to eliminate nonspecific reactivity due to protein A. Chu et al (6) recently reported the use of rabbit anti-protein A in a similar immunodot blot procedure for the detection of TSST-1. To further reduce the likelihood of cross-reactivity with protein A, the Fc portions of all primary and secondary detecting antibodies were preblocked with protein A.…”

Section: Discussionmentioning

confidence: 99%

Colony immunoblot assay for the detection of staphylococcal toxic shock syndrome toxin 1 (TSST-1) with anti-TSST-1 F(ab')2 fragments

et al. 1989

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Toxic shock syndrome toxin 1 (TSST-1), an exoprotein of Staphylococcus aureus, is strongly implicated in the pathogenesis of TSS. Detection of TSST-1, however, is often hindered in immunoassays because of the cosecretion of protein A, a genetic trait which appears to be coordinately expressed with other exoproteins in S. aureus. We developed a colony immunoblot assay for rapid screening of TSST-1-producing S. aureus using TSST-1-specific rabbit F(ab')2 fragments. The sensitivity and specificity of this method were compared with those of a quantitative noncompetitive enzyme-linked immunosorbent assay (ELISA) for 34 S. aureus isolates (17 TSS-associated and 17 non-TSS-associated isolates). Cosecreted protein A in culture supernatants was evaluated by a quantitative competitive ELISA. The results clearly indicated the superiority of F(ab')2 fragments in eliminating nonspecific reactivity of protein A in the colony immunoblot assay. The sensitivity of the immunoblot with TSST-1-specific F(ab')2 was similar to that with whole immunoglobulin G (94 versus 82%, respectively; P = 0.601, Fisher's exact test), but the specificity was markedly improved (94 versus 59%, respectively; P = 0.039). Among TSST-1-negative isolates (as determined by ELISA), strains which gave false-positive results in the immunoglobulin G immunoblot assay produced higher amounts of protein A than strains which gave true-negative results (P = 0.08, Mann-Whitney rank sum test, one tailed). Among strains positive for TSST-1, the level of TSST-1 detected in culture supernatants correlated inversely with the amount of protein A secreted (rs = -0.64; P less than 0.01, Spearman rank correlation). These findings validate the utility of a rapid screening method for the detection of TSST-1-producing S. aureus and support the concept of coordinate secretion of exoproteins in S. aureus.

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Association of toxic shock toxin-1 determinant with a heterologous insertion at multiple loci in the Staphylococcus aureus chromosome

Cited by 36 publications

References 40 publications

Cryptic α‐toxin gene in toxic shock syndrome and septicaemia strains of Staphylococcus aureus

Cryptic α‐toxin gene in toxic shock syndrome and septicaemia strains of Staphylococcus aureus

The gene for toxic shock toxin is carried by a family of mobile pathogenicity islands in Staphylococcus aureus

Colony immunoblot assay for the detection of staphylococcal toxic shock syndrome toxin 1 (TSST-1) with anti-TSST-1 F(ab')2 fragments

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