Molecular Detection of Antimicrobial Resistance

Fluit, Ad C.; Visser, Maarten R.; Schmitz, Franz‐Josef

doi:10.1128/cmr.14.4.836-871.2001

Cited by 401 publications

(301 citation statements)

References 405 publications

Supporting

Mentioning

277

Contrasting

Unclassified

Order By: Relevance

“…Mutations leading to quinolone resistance most frequently occur in the gene gyrA, namely in the quinolone resistance-determining region. At a chromosomal level, the development of resistance may be supported by reduced expression of outer membrane proteins or overexpression of efflux pumps (3,10).…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Quinolone-resistant Escherichia coli in Poultry Farming

Hricová¹,

Röderová²,

Pudová³

et al. 2017

Cent Eur J Public Health

View full text Add to dashboard Cite

SUMMARYIncreasing bacterial resistance to quinolone antibiotics is apparent in both humans and animals. For humans, a potential source of resistant bacteria may be animals or their products entering the human food chain, for example poultry. Between July 2013 and September 2014, samples were collected and analyzed in the Moravian regions of the Czech Republic to isolate the bacterium Escherichia coli. As a result, 212 E. coli isolates were obtained comprising 126 environmental isolates from poultry houses and 86 isolates from cloacal swabs from market-weight turkeys. Subsequently, the E. coli isolates were tested for susceptibility to selected antibiotics. Resistance of the poultry isolates to quinolones ranged from 53% to 73%. Additionally, the presence of plasmid-mediated resistance genes was studied. The genes were confirmed in 58% of the tested strains. The data on resistance of isolates from poultry were compared with results of resistance tests in human isolates obtained in the same regions.The high levels of resistance determined by both phenotyping and genotyping methods and reported in the present study confirm the fact that the use of fluoroquinolones in poultry should be closely monitored.

show abstract

Section: Introductionmentioning

confidence: 99%

“…DNA gyrase and topoisomerase IV are heterotetramers containing two subunits A and B. They are encoded by the genes gyrA and gyrB (in DNA gyrase) and parC and parE (in topoisomerase IV) (10,11). The development of bacterial resistance to quinolone antibiotics is a multifactorial process of both chromosomal and plasmid origin (12).…”

Section: Introductionmentioning

confidence: 99%

Quinolone-resistant Escherichia coli in Poultry Farming

Hricová¹,

Röderová²,

Pudová³

et al. 2017

Cent Eur J Public Health

View full text Add to dashboard Cite

show abstract

“…Various molecular methods have been used to identify the mutations in rpoB, katG, rpsL, rrs, embB, pncA, gyrA, and other genes (7,17). Among these methods, DNA sequencing is the most direct and reliable for detection of both known and novel mutations.…”

mentioning

confidence: 99%

Detection of Multidrug Resistance in Mycobacterium tuberculosis

et al. 2007

View full text Add to dashboard Cite

We developed a DNA sequencing-based method to detect mutations in the genome of drug-resistant Mycobacterium tuberculosis. Drug resistance in M. tuberculosis is caused by mutations in restricted regions of the genome. Eight genome regions associated with drug resistance, including rpoB for rifampin (RIF), katG and the mabA (fabG1)-inhA promoter for isoniazid (INH), embB for ethambutol (EMB), pncA for pyrazinamide (PZA), rpsL and rrs for streptomycin (STR), and gyrA for levofloxacin, were amplified simultaneously by PCR, and the DNA sequences were determined. It took 6.5 h to complete all procedures. Among the 138 clinical isolates tested, 55 were resistant to at least one drug. Thirty-four of 38 INH-resistant isolates (89.5%), 28 of 28 RIF-resistant isolates (100%), 15 of 18 EMB-resistant isolates (83.3%), 18 of 30 STR-resistant isolates (60%), and 17 of 17 PZA-resistant isolates (100%) had mutations related to specific drug resistance. Eighteen of these mutations had not been reported previously. These novel mutations include one in rpoB, eight in katG, one in the mabA-inhA regulatory region, two in embB, five in pncA, and one in rrs. Escherichia coli isolates expressing individually five of the eight katG mutations showed loss of catalase and INH oxidation activities, and isolates carrying any of the five pncA mutations showed no pyrazinamidase activity, indicating that these mutations are associated with INH and PZA resistance, respectively. Our sequencing-based method was also useful for testing sputa from tuberculosis patients and for screening of mutations in Mycobacterium bovis. In conclusion, our new method is useful for rapid detection of multiple-drug-resistant M. tuberculosis and for identifying novel mutations in drug-resistant M. tuberculosis.The emergence and spread of drug-resistant strains of Mycobacterium tuberculosis, especially multidrug-resistant (MDR) strains, are serious threats to the control of tuberculosis and comprise an increasing public health problem (40). Patients infected with MDR strains, which are defined as strains resistant to both rifampin (RIF) and isoniazid (INH), are difficult to cure and are more likely to remain sources of infection for a longer period of time than are patients with drug-susceptible strains (40).It is essential that rapid drug susceptibility tests be developed to prevent the spread of MDR M. tuberculosis. The time necessary for culture of specimens was reduced by the radiometric BACTEC 460TB system (BD Biosciences, Sparks, MD), the nonradiometric ESP II system (Trek Diagnostics, Westlake, OH), and other rapid broth methods, such as BACTEC MGIT 960 SIRE (BD Biosciences) (20). These drug susceptibility tests, however, still require 1 to 2 weeks for final determination and reporting to the clinician (23). Additional reductions in the detection period are needed.Drug resistance in M. tuberculosis is caused by mutations in relatively restricted regions of the genome (17, 39). Mutations associated with drug resistance occur in rpoB for RIF, katG and the promote...

show abstract

“…For example, β-lactamase production among members of the Enterobacteriaceae is common, but the development of resistance is dependent on the mode and level of expression. Molecular testing could however be required not only for therapy but also to monitor the spread of resistant organisms or resistance genes throughout the hospital and community (1).…”

mentioning

confidence: 99%

Clinical microbiologists, could we forget the history?

Migliavacca

Sambri

2017

Microbiol Med

View full text Add to dashboard Cite

SummaryClinical Microbiologists − whether they are medical doctors or biologists with postgraduate qualification in microbiology and virology − work in healthcare settings undertaking a wide range of laboratory analysis, monitoring microbial cultures and checking new drugs effects. Managing the laboratories, they ensure that data are recorded accurately, in agreement to the most updated clinical and analytical guidelines.Records analysis and interpretation are only the final steps of a complex processing work, whose quality depends on the choice of the most appropriate test and tool (and technical expertise), in agreement with the samples processing priorities.The Microbiology Technician figure represents another vital and crucial player in any Clinical Laboratory. This important professional figure is directly responsible for proper handling of all testing samples, any mishandling being able to end in poor testing and consequent inaccurate results.Microbiology Technicians are involved in setting up all the daily needed laboratory tests for microbial examination and, even before, they have to recognize if the specimens are well suitable for analytical processing. As well, sometimes it is required for these professional personnel to know how properly to store samples and/or strains, for further (confirmatory, deeper, molecular and/or epidemiological) investigations. Most of the microbiological identification work is performed using high-tech equipment, and today's Technicians must be proficient in computer programs to track the complete diagnostic pathway that specimens are following and to find related orders for testing (2).Different unsolved questions are however emerging, in the present era of microarray-based multiplexing and nucleic-acid-based deep-sequencing methods, which allow simultaneous detection of pathogen nucleic acid and multiple antibiotic resistance (4).As an example, does the acquisition of the most advanced molecular tools always positively affect the quality of the results and the laboratory productivity?Could the availability of high throughput instrumentation overcome a wasting time pre-analytical phase?In which cases do a molecular tool represent a priority? It is well known that MALDI-TOF mass spectrometry allowed us to achieve a more rapid and accurate germ identification results, making costs and work time at the bench lighter (3). Consequently, coupling to MALDI-TOF MS a mecA PCR targeting, positive blood cultures can now be reported up to 1.4 days earlier than it was previously possible, significantly improving the prognosis of MRSA caused bacteraemia by the administration of anti-MRSA active drugs within 48 hours after positive sample detection.An accurate and rapid diagnosis of infectious diseases is essential for appropriate antimicrobial use, but when will we be ready to discard in vitro bacterial growing tests?Despite the availability of molecular systems detecting antimicrobial resistance determinants (e.g. Extended Spectrum β-lactamases and/or carbapenemases genes), we cannot y...

show abstract

Molecular Detection of Antimicrobial Resistance

Cited by 401 publications

References 405 publications

Quinolone-resistant Escherichia coli in Poultry Farming

Quinolone-resistant Escherichia coli in Poultry Farming

Detection of Multidrug Resistance in Mycobacterium tuberculosis

Clinical microbiologists, could we forget the history?

Contact Info

Product

Resources

About