Mrsa screen

Manufactured by Denka Seiken

Sourced in Japan

The MRSA-Screen is a diagnostic test kit designed to detect the presence of Methicillin-resistant Staphylococcus aureus (MRSA) bacteria. It is a rapid and reliable screening tool for identifying MRSA colonization or infection.

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Lab products found in correlation

Slidex staph plus, by bioMérieux (2 mentions) Vitek 2 automated microbiology system, by bioMérieux (1 mentions) Iq sybr green supermix, by Bio-Rad (1 mentions)

4 protocols using mrsa screen

Methicillin Resistance Detection in S. pseudintermedius

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Methicillin resistance of isolated S. pseudintermedius was identified in two ways: a Kirby–Bauer disc diffusion test with an oxacillin disc (1 μg; Oxoid, Hampshire, UK), and a PCR assay targeting the mecA gene, as described previously (Zhang et al. 2005 ). Quality control of the disc diffusion test was performed using MR (ATCC BAA-44) and MS (ATCC 6538) strains of S. aureus. In the oxacillin disc diffusion test, a zone diameter ≤17 mm indicated a resistant strain (CLSI 2013 ).
When the results from the oxacillin disc diffusion test and the PCR assay detecting the mecA gene were discordant (i.e. a sample that yielded a mecA PCR amplicon but had a zone diameter ≥18 mm), a PBP2a latex agglutination test (MRSA-Screen; Denka Seiken, Tokyo, Japan) was additionally performed according to the manufacturer's protocol. If the isolate was found to be MR according to both the PCR and the PBP2a latex agglutination test, the isolate was considered a resistant strain.

Han J.I., Rhim H., Yang C.H, & Park H.M. (2017). Molecular characteristics of new clonal complexes of Staphylococcus pseudintermedius from clinically normal dogs. The Veterinary Quarterly, 38(1), 14-20.

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Identification of Methicillin-Resistant Staphylococcus aureus

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Samples were incubated in brain heart infusion (BHI) with 7% NaCl for 18–24 h at 37 °C. Then 10 µL of the infusion was plated onto MRSA-ID culture plate (bioMérieux^®, Durham, NC, USA). MRSA colonies were preliminarily identified as characteristically green malachite coloured, round colonies. Isolates were confirmed as S. aureus by Gram stain appearance, catalase test and coagulase test agglutination Slidex Staph Plus (bioMérieux^®). Species identification was confirmed by Vitek^® 2 Automated Microbiology System with the ID card GP (bioMérieux^®). S. aureus ATCC 29213 was used as the reference strain. Methicillin resistance was confirmed by testing the presence of penicillin-binding-protein A (PBP2a) (MRSA-screen; Denka Seiken Co™, Tokyo, Japan) and detecting the presence of the gene mecA by Real Time PCR (IQ™5; Bio-Rad, Hercules, CA, USA) [5 (link)].

Morcillo A., Castro B., Rodríguez-Álvarez C., Abreu R., Aguirre-Jaime A, & Arias A. (2015). Descriptive Analysis of Antibiotic-Resistant Patterns of Methicillin-Resistant Staphylococcus aureus (MRSA) st398 Isolated from Healthy Swine. International Journal of Environmental Research and Public Health, 12(1), 611-622.

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Rapid MRSA Identification via PBP2 Agglutination

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The MRSA-Screen latex agglutination test (MRSA-Screen; Denka Seiken Co, Ltd Tokyo, Japan) – a qualitative slide latex agglutination test for the detection of penicillin-binding protein 2′ (PBP2; also called PBP2a) present in isolates of S. aureus was performed according to the manufacturer’s instructions. The test is extremely useful in identifying MRSA. A suspension of 5 μL S. aureus organisms suspended in alkaline extraction agent (0.1 M NaOH) was boiled for 3 minutes, and 50 μL (1 drop) of extraction reagent 2 (0.5 M KH₂PO₄) was added. The mixture was centrifuged at 1,500×g for 5 minutes at room temperature, and 50 μL of the supernatant was mixed with the latex reagent on a test card. As a negative control, 50 μL of the supernatant was mixed with 1 drop (25 μL) of negative-control latex. After the contents of the slide were mixed on a shaker, agglutination within 3 minutes indicates the presumptive presence of_PBP2.31 (link)

Mocan L., Ilie I., Matea C., Tabaran F., Kalman E., Iancu C, & Mocan T. (2014). Surface plasmon resonance-induced photoactivation of gold nanoparticles as bactericidal agents against methicillin-resistant Staphylococcus aureus. International Journal of Nanomedicine, 9, 1453-1461.

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Methicillin-Resistant Staphylococcus aureus Identification

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Nasal swabs were incubated in brain heart infusion broth (BHI) with 7% NaCl for 18–24 h at 37 °C. Then, 10 µL of this broth was plated onto selective chromID^®MRSA agar culture (bioMérieux^®, Marcy l’Etoile, France), and incubated for 48 h at 37 °C. MRSA colonies were preliminarily identified as characteristic green malachite colored round colonies and purified on Columbia agar plates with 5% sheep blood (bioMérieux^®). Isolates were confirmed as S. aureus by Gram stain appearance catalase test and coagulase test agglutination (Slidex^® Staph Plus; bioMérieux^®). Species’ identifications were confirmed by Vitek^®2 Automated Microbiology System with a Gram positive identification card (bioMérieux^®). Methicillin resistance was confirmed by testing the presence of penicillin-binding protein A (PBP2a) (MRSA-screen; Denka Seiken Co™, Tokyo, Japan) and detecting the presence of the mecA gene by Real Time PCR (IQ™5; Bio-Rad, Hercules, CA, USA). The mecA gene (496bp) was amplified with the following primers: mecA- 1: 5′-GGG GTG GTT ACA ACG TTA CAA G-3′ mecA- 2: 5′-AGT TCT GCA GTA CCG GAT TTG C-3′. S. aureus ATCC 29213 was used as the reference strain. The primers were marked with IQ SYBR Green Supermix (BioRad^®, Madrid, Spain).

Abreu R., Rodríguez-Álvarez C., Lecuona M., Castro B., González J.C., Aguirre-Jaime A, & Arias Á. (2019). Increased Antimicrobial Resistance of MRSA Strains Isolated from Pigs in Spain between 2009 and 2018. Veterinary Sciences, 6(2), 38.

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