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Chapter 29 : Molecular Detection and Identification of Methicillin-Resistant

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Abstract:

This review focuses on the detection and identification of methicillin-susceptible (MRSA) both from primary microbiological (enrichment) cultures and clinical materials. The current gold standard method to identify from cultures is the AccuProbe Culture Identification Test (Gen-Probe). A major problem in classical MRSA diagnosis is the variable phenotypic expression of the gene-dependent methicillin resistance. With the availability of complete inventories of putative virulence genes, based on whole-genome comparisons, the possibilities for targeted diagnosis will increase in the future. The Velogene Rapid MRSA identification assay is based on a chimeric probe targeting the gene. Pooling of clinical swabs and process automation will reduce the costs in the future. It needs to be emphasized that in principle the issues covered in this review can be extrapolated to species and isolates of each and every other antibiotic-resistant microbial infectious disease agent including, for instance, vancomycin-resistant enterococci. Molecular technology has changed the horizon, and for , for instance, molecular detection already is the gold standard technology. That molecular testing will also revolutionize MRSA detection is obvious. It remains to be seen which of the many currently available technologies will in the end be the one that is collectively embraced by the majority of clinical microbiologists.

Citation: Leeuwen W, Belkum A. 2011. Molecular Detection and Identification of Methicillin-Resistant , p 463-477. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch29

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Multiplex Real-Time PCR
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Fourier Transform Infrared Spectroscopy
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Mobile Genetic Elements
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FIGURE 1

Organization of the known SCC types. SCC elements share four characteristics: (i) the gene complex (dotted boxes) consisting of the methicillin resistance determinant, presence or absence of (parts of) its regulatory genes, and insertion sequences (IS); (ii) presence of the cassette chromosome recombinase genes responsible for the mobility of the SCC element; (iii) presence of direct and inverted complementary repeat sequences at both ends of the element; and (iv) integration of the element on the staphylococcal chromosome into the 3′-end of open reading frame X . SCC type definition is based on the identification of its components: genes (five types), complex (four classes), and specific structures in junkyard (J) regions (plasmids and transposons). The subtypes (not indicated) within the SCC types II and III are characterized by junkyard sequence variability. The approximate positions of the forward (▶) and reverse (◀) primers and detection probes (bars) used in the Huletsky PCR strategy for MRSA detection and identification are shown. The structures of the SCC elements are based on the nucleotide sequences coded for GenBank accession nos. AB033763 (type I), D86934 (type II), AB037671 (type III), AB063172 (type IV), AB121219 (type V), and AF411935 (typeVI). The SCC element typing nomenclature is based on the work of Kondo et al. ( ).

Citation: Leeuwen W, Belkum A. 2011. Molecular Detection and Identification of Methicillin-Resistant , p 463-477. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch29
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Tables

Generic image for table
TABLE 1

Commercial diagnostic tests for the identification of

Citation: Leeuwen W, Belkum A. 2011. Molecular Detection and Identification of Methicillin-Resistant , p 463-477. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch29
Generic image for table
TABLE 2

Evaluation studies of selective agar plates for direct MRSA screening in swabs

Citation: Leeuwen W, Belkum A. 2011. Molecular Detection and Identification of Methicillin-Resistant , p 463-477. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch29
Generic image for table
TABLE 3

Commercial immunological tests for MRSA identification and molecular tests for MRSA detection and identification

Citation: Leeuwen W, Belkum A. 2011. Molecular Detection and Identification of Methicillin-Resistant , p 463-477. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch29
Generic image for table
TABLE 4

In-house molecular tests for MRSA detection and identification

Citation: Leeuwen W, Belkum A. 2011. Molecular Detection and Identification of Methicillin-Resistant , p 463-477. In Persing D, Tenover F, Tang Y, Nolte F, Hayden R, van Belkum A (ed), Molecular Microbiology. ASM Press, Washington, DC. doi: 10.1128/9781555816834.ch29

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