[The microbiology laboratory's contribution to the surveillance and control of outbreaks caused by nonfermentative Gram-negative bacilli].
Unidad Clínica de Enfermedades Infecciosas y Microbiología, Hospital Universitario Virgen Macarena, Sevilla, España.Enfermedades Infecciosas y Microbiología Clínica (Impact Factor: 2.17). 03/2011; 29 Suppl 3:40-6.
Pseudomonas aeruginosa, Acinetobacter baumannii and Stenotrophomonas maltophilia are, among others, the most important nonfermentative Gram-negative bacilli within hospitals. These organisms are able to cause different types of nosocomial infections in predisposed patients, and P. aeruginosa and A. baumannii in particular may cause extensive clonal outbreaks and complex situations in which sporadic isolates may coexist with some epidemic strains. Some common features are their intrinsic resistance to many antimicrobials, their capacity to further develop antimicrobial resistance, and the possibility of environmental reservoirs as sources of these microorganisms in healthcare centers. The microbiology laboratory plays a key role in the detection of potential outbreaks, in the identification of new resistance mechanisms and in the characterization of local epidemiology, by detecting colonized patients and/or environmental reservoirs if needed, appropriately identifying the isolates, phenotypically or genotypically characterizing their mechanisms of resistance, if appropriate, and finally by using different molecular techniques for clonal typing, which are reviewed in this article. Such activities must be performed in the context of the surveillance and control programs of specific institutions and as part of the daily work of multidisciplinary infection control teams.
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ABSTRACT: Molecular typing methods are useful in the surveillance and control of nosocomial outbreaks because they can provide information on the clonal relatedness among isolates, identify reservoirs, and determine routes of transmission. The gold standard assay for molecular typing is pulsed-field gel electrophoresis (PFGE) due to its high discriminatory power. Some major disadvantages of PFGE include the high cost of the equipment, its labor intensiveness (the technique is not automated) and the time required to analyze the profiles of DNA bands (pulsotypes). Although there are many molecular typing methods based on polymerase-chain reaction (PCR), the most widely used is repetitive sequence-based PCR (REP-PCR). Most of the PCR techniques used for molecular typing have none of the limitations of PFGE as they are less expensive and labor intensive (some, such as bioMérieux's Diversilab system, are commercially available) and generate DNA profiles that are easier to interpret, depending on the microorganism. The discriminatory power of PCR is generally lower than or similar to that of PFGE. Both PFGE and PCR require optimal laboratory standardization to guarantee good reproducibility. PCR methods are preferable in the study of small, timelimited outbreaks. In more complex outbreaks of longer duration, in which clonal evolution and dynamics are studied, the use of PFGE is preferable. Molecular typing methods based on DNA sequencing, such as multilocus sequence typing, are applicable in global epidemiological studies or in analyses of the population structure of microorganisms.
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ABSTRACT: Healthcare-associated infections due to multidrug-resistant Gram-negative bacteria (MDR-GNB) are a leading cause of morbidity and mortality worldwide. These evidence-based guidelines have been produced after a systematic review of published studies on infection prevention and control interventions aimed at reducing the transmission of MDR-GNB. The recommendations are stratified by type of infection prevention and control intervention and species of MDR-GNB and are presented in the form of 'basic' practices, recommended for all acute care facilities, and 'additional special approaches' to be considered when there is still clinical and/or epidemiological and/or molecular evidence of ongoing transmission, despite the application of the basic measures. The level of evidence for and strength of each recommendation, were defined according to the GRADE approach.
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