Real-time PCR for universal antibiotic susceptibility testing

Rolain, Jean‐Marc; Mallet, M N; Fournier, Pierre‐Edouard; Raoult, Didier

doi:10.1093/jac/dkh324

Cited by 91 publications

(63 citation statements)

References 9 publications

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“…The MICs determined using real-time PCR were almost similar to those obtained by conventional methods. Rolain et al (29) showed that it was possible to speed up the susceptibility tests also for commonly isolated bacteria like Staphylococcus aureus, Haemophilus influenzae, Escherichia coli, and others by real-time PCR. The results were in accordance with those obtained following the CLSI (formerly NCCLS) guidelines.…”

Section: Discussionmentioning

confidence: 99%

Antibiotic Susceptibility Testing of Mycoplasma genitalium by TaqMan 5′ Nuclease Real-Time PCR

Hamasuna

Osada

Jensen

2005

Antimicrob Agents Chemother

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Mycoplasma genitalium is an important pathogen in male nongonococcal urethritis (NGU). Isolation of M. genitalium from clinical specimens by axenic culture is very difficult and time-consuming, and very few strains are available for antibiotic susceptibility testing. Primary isolation of M. genitalium by coculture with Vero cells improves the isolation rate significantly. However, some strains cannot be adapted to axenic culture. In this study, we determined the antibiotic susceptibility of M. genitalium strains grown in Vero cell culture with dilutions of antibiotics. Growth of M. genitalium was monitored by a quantitative PCR assay detecting a single-copy region of the mgpB adhesin gene. Growth inhibition in the presence of antibiotics was expressed as a percentage of the DNA load of controls grown in the absence of antibiotics. Eighteen strains were examined, including 6 new strains isolated from urethral swab specimens and 4 new strains isolated from urine specimens collected from Japanese men. Eight strains adapted to axenic culture were also tested by the conventional broth dilution method. The two methods had an acceptable correlation. Azithromycin was the most active drug against M. genitalium. Among the fluoroquinolones, moxifloxacin had the highest activity, with MICs ranging from 0.03 to 0.5 mg/liter, whereas ciprofloxacin and levofloxacin were considerably less active, with MICs ranging from 0.5 to 16 mg/liter and 0.25 to 4 mg/liter, respectively. MICs for tetracycline ranged from 0.125 to 4 mg/liter. This new method could increase the number of M. genitalium strains available for antibiotic susceptibility testing and significantly shorten the time from sampling to MIC results.Mycoplasma genitalium was first isolated after prolonged incubation from 2 of 13 men with urethritis (36). Despite repeated attempts, new isolates of M. genitalium were not obtained from urethral specimens by conventional culture techniques (31, 33), but isolation of M. genitalium in cultures of throat and synovial fluid specimens has been reported, although these isolates were mixed with Mycoplasma pneumoniae (1, 35). Aside from these reports, a technique for isolation of M. genitalium from urethral specimens using Vero cells to initially cultivate M. genitalium and subsequent adaptation to conventional broth has been developed (19) and has been used with some success in other laboratories (34). Using this new technique, however, it took more than 50 days to adapt the M. genitalium strains to cell-free medium, i.e., axenic culture. Even today, other urogenital isolates are extremely rare and have been difficult to obtain (23).M. genitalium is gaining increasing attention as one of the important pathogens in men with nongonococcal urethritis (NGU). While Chlamydia trachomatis can be found in 30 to 40% of NGU patients, M. genitalium has been detected in 15 to 30% (3,11,15,20). Several studies have shown a significant association between the presence of M. genitalium DNA and symptoms and/or signs of urethritis (reviewed in refere...

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Section: Discussionmentioning

confidence: 99%

Antibiotic Susceptibility Testing of Mycoplasma genitalium by TaqMan 5′ Nuclease Real-Time PCR

Hamasuna

Osada

Jensen

2005

Antimicrob Agents Chemother

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“…The potential exists for molecular methods, such as a multiplex PCR approach or DNA microarray technology, to simultaneously identify the presence of multiple resistance genes in just a few hours (93). Alternatively to the detection of resistance genes, a universal 16S rRNA gene-and rpoB-based real-time quantitative PCR assay for susceptibility testing of bacteria has been devised to decrease the time needed for antimicrobial testing results (229).…”

Section: Future Directionsmentioning

confidence: 99%

Microbiology and Treatment of Acute Apical Abscesses

2013

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SUMMARY Acute apical abscess is the most common form of dental abscess and is caused by infection of the root canal of the tooth. It is usually localized intraorally, but in some cases the apical abscess may spread and result in severe complications or even mortality. The reasons why dental root canal infections can become symptomatic and evolve to severe spreading and sometimes life-threatening abscesses remain elusive. Studies using culture and advanced molecular microbiology methods for microbial identification in apical abscesses have demonstrated a multispecies community conspicuously dominated by anaerobic bacteria. Species/phylotypes commonly found in these infections belong to the genera Fusobacterium , Parvimonas , Prevotella , Porphyromonas , Dialister , Streptococcus , and Treponema . Advances in DNA sequencing technologies and computational biology have substantially enhanced the knowledge of the microbiota associated with acute apical abscesses and shed some light on the etiopathogeny of this disease. Species richness and abundance and the resulting network of interactions among community members may affect the collective pathogenicity and contribute to the development of acute infections. Disease modifiers, including transient or permanent host-related factors, may also influence the development and severity of acute abscesses. This review focuses on the current evidence about the etiology and treatment of acute apical abscesses and how the process is influenced by host-related factors and proposes future directions in research, diagnosis, and therapeutic approaches to deal with this disease.

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“…As infectious disease testing is by far the most popular area for use of NADs we will focus our attention on this application. Within the field of infectious disease testing, key areas where NADs are employed include microbial identification, detection of pathogen antibiotic susceptibility [42] and monitoring and surveillance of routes of infection [43]. The use of NADs in these areas has provided rapid sensitive tools for pathogen detection and identification, and for establishing antibiotic susceptibility, ultimately leading to improved treatment options.…”

Section: Use Of Nads In Clinical Practice and Disease Managementmentioning

confidence: 99%