Serum antibodies to a Staphylococcus aureus thermonuclease preparation in healthy persons and patients with bacteremia

Brakstad, Odd Gunnar; Mæland, Johan A.; Wergeland, Heidrun I.

doi:10.1016/0888-0786(89)90034-6

Cited by 9 publications

(3 citation statements)

References 20 publications

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“…One of the strategies to identify MRSA is multiplex PCR, developed for simultaneous amplification of methicillin resistance gene, mecA and one of the S. aureus-specific genes, such as coa, gyrA, holB (SA442), femA, femB or nuc gene, encoding for S. aureus specific thermonuclease. In a comparative study, Brakstad et al (1989) used a multiplex PCR targeting mec A and nuc which report a 100% agreement with conventional identification methods. This approach is generally applied for the identification of subcultures of MRSA in routine diagnostic microbiological laboratory.…”

Section: Discussionmentioning

confidence: 99%

Study on prevalence and genetic discrimination of methicillin-resistant Staphylococcus aureus (MRSA) in Egyptian hospitals

Elshimy¹,

Khattab²,

Zedan³

et al. 2018

Afr. J. Microbiol. Res.

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Methicillin-resistantStaphylococcus aureus (MRSA) continues to be a global problem in infection control. The highest proportions of MRSA are reported by Jordan, Egypt and Cyprus investigators, where more than 50% of the invasive isolates are methicillin-resistant. The aim of this work was to study the prevalence, antibiotic sensitivity and genetic discrimination of MRSA in Egypt. Microbiological identification was done using Gram stain, catalase, coagulase and mannitol fermentation along with biochemical identification by analytical profile index (API) tests. Molecular identification was conducted by the polymerase chain reaction (PCR) targeting 16S ribosomal RNA and the nuc genes. Additionally, identification of methicillin-resistant S. aureus (MRSA) was performed by the amplification of 310 bp of the mecA gene. Antibiograms were performed for all isolates. Only 73 isolates out of 166 were oxacillin resistant. The percentage of resistant isolates to erythromycin, rifampicin, vancomycin, Ofloxacin, gentamycin, Amoxicillin clavulanic acid, ciprofloxacin, chloramphenicol, trimethoprim sulfamethoxazole, teicoplanin and tetracycline were 58, 32.50, 2.4, 45.18, 37.9, 39.7, 23.5, 21.6, 40.3, 0 and 39.1%, respectively. MRSA isolates were subdivided into eight biotypes according to their resistance pattern. Random amplification of polymorphic DNA (RAPD) and repetitive sequence DNA (REP) were performed on samples representing each biotype.

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Section: Discussionmentioning

confidence: 99%

Study on prevalence and genetic discrimination of methicillin-resistant Staphylococcus aureus (MRSA) in Egyptian hospitals

Elshimy¹,

Khattab²,

Zedan³

et al. 2018

Afr. J. Microbiol. Res.

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“…Heating of the blood culture samples before testing by the MAb-based methods was necessary in order to detect the enzyme protein, probably due to immune complex formation with anti-TNase antibodies in human blood (5). Experiments also showed that the TNase was not bound by the resin.…”

Section: Discussionmentioning

confidence: 99%

“…Products detected by gel electrophoresis (A) and sELNFA (B) after modified nuc-PCR( 1 4 ) or conventional nuc-PCR(5)(6)(7)(8). Dilutions of the S. aureus strain 50020/92 prepared in blood culture medium supplemented with blood were tested.…”

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confidence: 99%

Direct identification of Staphylococcus aureus in blood cultures by detection of the gene encoding the thermostable nuclease or the gene product

Brakstad

Mæland²

1995

APMIS

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This study compares methods for direct identification of S. aureus in blood cultures by detection of the thermonuclease (TNase) of this bacterium or the nuc gene encoding it. The protein was detected by an enzyme diffusion test in o‐toluidine blue DNA agar with a test time of at least 4 h, by a monoclonal antibody (MAb)‐based sandwich enzyme‐linked immunosorbent assay (sELISA) with a test time of ˜4 h, and by a MAb‐based sandwich enzyme‐linked immunofiltration assay (sELIFA) with a test time of 25–30 min (sample preparation included). The nuc gene was amplified by a polymerase chain reaction (PCR) with a test time (amplification plus detection) of ‐3.5 h. The tests were optimized for direct examination of blood‐containing cultures. All tests were positive with 67/67 blood cultures which grew S. aureus, negative with 35/35 cultures which grew coagulase‐negative staphylococci, and negative with 37/37 cultures with various other bacteria. These results showed positive agreement with those of the commercial AccuProbe test but not with the StaphAurex agglutination kit. With an artificially seeded blood culture, minimum total times required (incubation plus testing) were as follows: nuc‐PCR, 9.5 h; sELIFA, 12.5 h; enzymatic test, 16–36 h; AccuProbe, 14 h. Direct examination of both the nuc gene and the mecA gene encoding methicillin resistance demonstrated the mecA gene in all the coagulase‐negative staphylococci (48.6%) which showed oxacillin resistance. The sELIFA had the particular advantage of its short test time, the PCR its high sensitivity and the possibility of simultaneous detection of the species‐specific nuc gene and genes encoding other clinically important characters of the bacteria. These tests offer the prospect of direct application to a variety of clinical specimens for rapid diagnosis of S. aureus infection.

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Use of Immunodiagnostics for the Early Detection of Biofilm Infections

Selan¹,

Kofonow²,

Scoarughi³

et al. 2008

Springer Series on Biofilms

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Serum antibodies to a Staphylococcus aureus thermonuclease preparation in healthy persons and patients with bacteremia

Cited by 9 publications

References 20 publications

Study on prevalence and genetic discrimination of methicillin-resistant Staphylococcus aureus (MRSA) in Egyptian hospitals

Study on prevalence and genetic discrimination of methicillin-resistant Staphylococcus aureus (MRSA) in Egyptian hospitals

Direct identification of Staphylococcus aureus in blood cultures by detection of the gene encoding the thermostable nuclease or the gene product

Use of Immunodiagnostics for the Early Detection of Biofilm Infections

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