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Chapter 4 : Molecular Methods for Detection of Antibiotic Resistance

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Abstract:

The combination of molecular characterization of resistant strains with precise identification of the antibiotic resistance gene(s) or mutation(s) and the genetic elements involved in the dissemination of these genes is an effective approach in the control of the spread of antibiotic resistance. Molecular methods also help to determine the location of the gene and to differentiate between horizontal gene transfer and clonal spread. This chapter describes polymerase chain reaction (PCR) and microarray analysis in detail, and specifies applications of these methods in the investigation of antibiotic-resistant strains. Numerous PCR assays for the detection of antibiotic resistance genes have been developed and the development of microarrays for the simultaneous detection of a large number of these genes and the genetic elements involved in their dissemination is in progress. The implementation of molecular methods in routine analysis can be achieved only when it is supported by the proper validation of the methods and the availability of the necessary controls, reference strains, and educated personnel. The molecular characterization of antibiotic-resistant strains can help to identify atypical resistant strains, describe new outbreak strains at an early stage, elucidate the epidemiology of resistant strains at a genotypic level, and explain the processes leading to the selection of resistant and virulent strains. In addition, molecular methods can allow proper risk assessment with respect to the use of antimicrobial substances. If the transcriptional and translational expression of antibiotic resistance genes can be better understood, molecular methods may replace phenotypic measurements.

Citation: Aarts H, Guerra B, Malorny B. 2006. Molecular Methods for Detection of Antibiotic Resistance, p 37-48. In Aarestrup F (ed), Antimicrobial Resistance in Bacteria of Animal Origin. ASM Press, Washington, DC. doi: 10.1128/9781555817534.ch4

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Mobile Genetic Elements
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Figure 1

Detection methods in real-time PCR. (A) SYBR Green emits a fluorescent signal when it binds to dsDNA. (B) Specific binding of a double-labeled hydrolysis, or TaqMan, probe. Q indicates the quencher dye and F the fluorescent (reporter) dye. (C) Specific binding of the hybridization FRET probes (acceptor and donor). The donor fluorescence (D) is transmitted to the acceptor fluorescence (AF) only when it is in close proximity, generating a fluorescence signal. See the text for further explanation.

Citation: Aarts H, Guerra B, Malorny B. 2006. Molecular Methods for Detection of Antibiotic Resistance, p 37-48. In Aarestrup F (ed), Antimicrobial Resistance in Bacteria of Animal Origin. ASM Press, Washington, DC. doi: 10.1128/9781555817534.ch4
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References

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Tables

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Table 1

Real-time PCR assays for the detection of antibiotic resistance genes

Citation: Aarts H, Guerra B, Malorny B. 2006. Molecular Methods for Detection of Antibiotic Resistance, p 37-48. In Aarestrup F (ed), Antimicrobial Resistance in Bacteria of Animal Origin. ASM Press, Washington, DC. doi: 10.1128/9781555817534.ch4

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