A new closed-tube multiplex real-time PCR to detect eleven superantigens of Streptococcus pyogenes identifies a strain without superantigen activity

Lintges, Maria; Arlt, Sabine; Uciechowski, Peter; Plümäkers, Birgit; Reinert, Ralf René; Al‐Lahham, Adnan; Lütticken, Rudolf; Rink, Lothar

doi:10.1016/j.ijmm.2007.03.015

Cited by 29 publications

(24 citation statements)

References 52 publications

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“…Molecular analysis of the emm genes was performed by PCR using "all M primers" (1) and resulted in the genotype emm-4. Profiling of the presence of streptococcal pyrogenic exotoxin genes (spe), the streptococcal mitogenic exotoxin Z gene (smeZ), and the streptococcal superantigen gene (ssa) was performed as described previously (2). This analysis revealed the coexistence of speC, smeZ, and ssa, in accordance with reports demonstrating the association of these superantigen genes with the emm-4 lineage (3, 4).…”

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confidence: 72%

Severe Soft Tissue Infection Caused by a Non-Beta-Hemolytic Streptococcus pyogenes Strain Harboring a Premature Stop Mutation in the sagC Gene

et al. 2013

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gWe recovered a non-beta-hemolytic Streptococcus pyogenes strain from a severe soft tissue infection. In this isolate, we detected a premature stop codon within the sagC gene of the streptolysin S (SLS) biosynthetic operon. Reintroduction of full-length sagC gene on a plasmid vector restored the beta-hemolytic phenotype to our clinical isolate, indicating that the point mutation in sagC accounted for loss of hemolytic activity. To the best of our knowledge, this is the first report to demonstrate that a severe soft tissue infection can be caused by a non-beta-hemolytic S. pyogenes strain lacking a functional SagC. CASE REPORTA 33-year-old man developed pain and swelling after an insect bite to his left forearm. Conservative therapy on an outpatient basis was initiated involving oral roxithromycin and splinting with a cast. The past medical history was uneventful except for a herniated intervertebral disc and arterial hypertension. Except for the prescribed antibiotic, the patient was on no other medication. After 1 week of treatment without any improvement, the patient was referred to the Department of Plastic and Hand Surgery with an abscess on the distal left forearm.On examination, the obese patient (body mass index [BMI], 32 kg/m 2 ) was afebrile. His blood pressure was 140/80 mm Hg, and his pulse was 80 beats per minute. According to the patient, there had previously been pronounced lymphangitis involving the forearm that had improved by the time of presentation to our department. Around the distal forearm, there was an abscess with central soft tissue necrosis (Fig. 1A). The surrounding soft tissue was erythematous, swollen, and tender (Fig. 1A). The sensation of light touch was intact, and the patient was able to move his fingers. There was no crepitation. Lab tests revealed a leukocytosis (11.17/ nl; normal range, 4 to 10/nl) and an elevated C-reactive protein level (52.4 mg/liter; normal range, Ͻ5 mg/liter).Emergency surgery with abscess incision and drainage was performed. Necrotic tissue was removed. Upon further surgical exploration, the peritendineous area surrounding the flexor carpi radialis and the palmaris longus tendons showed extensive pannus formation extending to and involving the loose tissue surrounding the radial artery, suggesting a longer duration of the infectious process prior to surgery. Tenolysis and microsurgical arteriolysis of the radial artery were performed meticulously, and surgical débridement of the abscess was achieved. Tissue specimens were sent for Gram staining and bacterial culture, followed by placement of drains for irrigation and suction of the wound bed.An antibiotic regimen with amoxicillin-clavulanic acid was initiated empirically and continued for 2 weeks. The patient fully recovered and was discharged. Since surgical débridement left only a small defect, no plastic surgery was necessary and the wound healed uneventfully.Direct examination of the surgical tissue specimen revealed abundant neutrophils and Gram-positive cocci ordered in chains. Cultures of surgical...

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Severe Soft Tissue Infection Caused by a Non-Beta-Hemolytic Streptococcus pyogenes Strain Harboring a Premature Stop Mutation in the sagC Gene

et al. 2013

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“…The strains were cultivated on sheep blood agar plates at 37ЊC in a 5% carbon dioxide environment. S. pyogenes was identified by b-hemolysis and further characterized using Lancefield antigen typing, as described elsewhere [30].…”

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confidence: 99%

“…Multiplex PCR was established and performed as described elsewhere [30]. Briefly, a multiplex PCR kit (Qiagen) was used along with 2 different primer mixes and genomic DNA.…”

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confidence: 99%

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Superantigen Genes Are More Important than theemmType for the Invasiveness of Group AStreptococcusInfection

et al. 2010

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These results indicate that the presence of superantigen genes is more important for the invasiveness of group A Streptococcus infection than emm type and may be the connection between the high-risk HLA type of the host and the pathogen. Furthermore, we found a very clear correlation between the presence of the genes spea1-spea3 and the presence of the gene emm1, which indicates that the relationship between emm1 and invasiveness is based on the superantigen gene profile. Our data suggest that the superantigen gene profile is of high importance for the clinical outcome of group A Streptococcus infections.

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“…1, this emm gene had a single nucleotide deletion in the hypervariable 5Ј end compared to other alleles of sequence type stG6 and was predicted to translate to a truncated M protein of only 56 residues, lacking the conserved domains and anchor region. A multiplex PCR with previously described primer pairs was conducted for detection of the 11 streptococcal superantigens speA, speC, speG, speH, speI, speJ, speK, speL, speM, smeZ, and ssa (17). In order to cover the allelic variation of the smeZ gene, we also used a single PCR with a previously reported primer pair (5).…”

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Two Unusual Cases of Severe Soft Tissue Infection Caused by Streptococcus dysgalactiae subsp. equisimilis

et al. 2010

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We present two cases of invasive infection caused by Streptococcus dysgalactiae subsp. equisimilis, one that showed rapidly developing necrotizing fasciitis in a previously healthy man and one that showed severe cellulitis and septic shock even though the bacterium possessed a mutated emm gene, predicted to encode a truncated M protein. CASE REPORTPatient 1 was a 48-year-old, previously healthy man admitted to Haukeland University Hospital in Western Norway with a fever and a rapidly spreading erythema of the skin overlying his right knee. The symptoms were preceded by a minor trauma toward the right knee 5 h prior to admission, leading to a disruption of the skin barrier. Blood pressure upon admission was 127/68 mm Hg, pulse rate 90/min, and temperature 39.6°C. Blood cultures were taken, and intravenous treatment with penicillin G at 5 million IU four times a day (q.i.d.) and clindamycin at 900 mg three times a day (t.i.d.) was started. Surgical exploration was performed 2 h later because of rapid deterioration of the patient's condition, and necrotic subcutaneous tissue and fascia of the lateral margin of the patella and the upper part of the calf were excised. Hypotension (blood pressure, 85/45 mm Hg) developed postoperatively, and normal blood pressure was reestablished after 5 to 6 h of intensive intravenous fluid therapy. Neither renal, hepatic, nor respiratory failure developed. The international normalized ratio (INR; normal range, Ͻ1

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A new closed-tube multiplex real-time PCR to detect eleven superantigens of Streptococcus pyogenes identifies a strain without superantigen activity

Cited by 29 publications

References 52 publications

Severe Soft Tissue Infection Caused by a Non-Beta-Hemolytic Streptococcus pyogenes Strain Harboring a Premature Stop Mutation in the sagC Gene

Severe Soft Tissue Infection Caused by a Non-Beta-Hemolytic Streptococcus pyogenes Strain Harboring a Premature Stop Mutation in the sagC Gene

Superantigen Genes Are More Important than theemmType for the Invasiveness of Group AStreptococcusInfection

Two Unusual Cases of Severe Soft Tissue Infection Caused by Streptococcus dysgalactiae subsp. equisimilis

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