Accuracy and Potential Usefulness of Triplex Real-Time PCR for Improving Antibiotic Treatment of Patients with Blood Cultures Showing Clustered Gram-Positive Cocci on Direct Smears

Ruimy, Raymond; Dos-Santos, Marie; Raskine, Laurent; Bert, Frédéric; Masson, René; Elbaz, Sandrine; Bonnal, Christine; Lucet, Jean‐Christophe; Lefort, A.; Fantin, Bruno; Wolff, Michel; Hornstein, M; Andremont, Antoine

doi:10.1128/jcm.02250-07

Cited by 56 publications

(43 citation statements)

References 45 publications

(54 reference statements)

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“…E. coli isolates were tested for susceptibility to amoxicillin, tircacillin, amoxicillin + clavulanic acid, cefotaxime, cefoxitin, ceftazidime, cefepime, ertapenem, nalidixic acid, ciprofloxacin, gentamicin, amikacin, fosfomycin and co-trimoxazole (Bio-Rad, Marne-la-Coquette, France) using the disc diffusion method and presence of ESBL was determined using the double disc-diffusion phenotypic method, as recommended by the Antibiogram Committee of the French Society for Microbiology [4]. To confirm the ESBL identification with molecular methods, total DNA was extracted from E. coli strains, as described [5] and blaCTX-M (group 1), blaTEM and blaSHV genes were amplified by PCR using specific primers, as described [6].…”

Section: Microbiological Analysismentioning

confidence: 99%

Intestinal carriage of Extended Spectrum Beta-Lactamase producing E. coli in women with urinary tract infections, Cameroon

Djuikoue

Woerther

Toukam

et al. 2016

J Infect Dev Ctries

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Introduction: During the last decade, the prevalence of the intestinal carriage of extended spectrum beta-lactamases -producing Escherichia coli (ESBL-E. coli) has continued to increase worldwide in the community, especially in developing countries. Hence, we undertook a study to determine the ESBL-E. coli fecal carriage rate and the associated risk factors in Cameroonian women. Methodology: A total of 86 women suspected of community-acquired urinary tract infections (UTI) were included in 10 health structures from May 2011 to April 2012. After filling a questionnaire, they provided a stool sample that was plated on selective media for ESBL producing bacteria. The identification of strains was obtained with mass spectrometry and the antibiotic susceptibility by disk diffusion in agar media. The ESBL type was determined by PCR. The relative abundance of ESBL-E. coli was measured for positive samples. Eventually, the presence of antibiotics in stool was assessed. Results: The carriage rate of ESBL-E. coli was 57/86 (66.3%). Phenotypic and molecular characterization showed that all ESBL-E. coli strains contained group 1 CTX-M enzymes. Multivariate analysis showed that ESBL-E. coli fecal carriage was associated with the presence of antibiotics in stools (p < 0.05). Although not significant, mean ESBL relative abundance tended to be higher in patients with antibiotic exposure. Conclusions: Our results show that the carriage of ESBL-E. coli fecal carriage in women with UTI suspicion from the Cameroonian community is extremely high and associated with recent antibiotic intake.

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Section: Microbiological Analysismentioning

confidence: 99%

Intestinal carriage of Extended Spectrum Beta-Lactamase producing E. coli in women with urinary tract infections, Cameroon

Djuikoue

Woerther

Toukam

et al. 2016

J Infect Dev Ctries

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“…Automated blood culture systems take approximately 1 to 2 days, on average, to signal a positive blood culture and another 1 to 2 days to finalize bacterial identification and antimicrobial testing. With the advent of qPCR, the time to bacterial identification and detection of drug resistance has been reduced to 4 to 6 h after a positive blood culture has turned positive (22,24,26,28). However, the presence of PCR inhibitors in the blood culture bottles has thus far reduced the sensitivity of PCR assays (10).…”

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confidence: 99%

Rapid Detection of bla _KPC Carbapenemase Genes by Internally Controlled Real-Time PCR Assay Using Bactec Blood Culture Bottles

et al. 2011

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Rapid detection of drug-resistant bacteria in clinical samples plays an instrumental role in patients'infection management and in implementing effective infection control policies. In the study described in this report, we validated a multiplex TaqMan real-time quantitative PCR (qPCR) assay for the detection of bla KPC genes and the human RNase P gene in Bactec blood culture bottles. The MagNA Pure LC (version 2.0) instrument was utilized to extract nucleic acids from the inoculated broth, while bovine serum albumin (BSA) was utilized as the PCR inhibitor reliever. The multiplex assay, which was specific for the detection of bla KPC genes, had a limit of detection of 19 CFU per reaction mixture with human blood-spiked Bactec bottles. Of the 323 Bactec blood culture sets evaluated, the same 55 (17%) blood cultures positive for carbapenem-resistant bacteria by culture were also positive by the validated qPCR assay. Thus, the sensitivity, specificity, positive predictive value, and negative predictive value of the qPCR assay compared to the results of culture were all 100%. bla KPC genes were also detected from the same Bactec bottle broth after manual extraction with a QIAamp DNA minikit; however, there was an average 3-threshold-cycle delay in the qPCR readings. With the limited therapeutic options available, the accurate and rapid detection of bla KPC -possessing bacteria by the described bla KPC /RNase P assay will be a crucial first step in ensuring optimal clinical outcomes and infection control.

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“…Three MSSA strains (13,30, and 114) were misidentified as MRSA by the BD GeneOhm StaphSR assay. None of these strains harbored a functional mecA gene as analyzed by PCR techniques, and no SCCmec type could be determined.…”

Section: Discussionmentioning

confidence: 99%

“…However, bacterial identification and preliminary antibiotic susceptibility testing by standard microbiological procedures still requires 24 to 48 h after growth detection by automated incubation systems. In contrast, new real-time PCR-based methods that use samples directly from positive blood culture bottles allows differentiation of MSSA, MRSA, and coagulase-negative staphylococci (CoNS) within 1.5 to 3 h (7,12,13,16). Such tests promote an early appropriate antibiotic selection, thus avoiding the unnecessary use of vancomycin, and they reduce mortality, the length of hospitalization, and costs associated with bloodstream infections caused by these bacteria (3).…”

mentioning

confidence: 99%

Evaluation of the BD GeneOhm StaphSR Assay for Detection of Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Isolates from Spiked Positive Blood Culture Bottles

Gröbner

Dion²,

Plante³

et al. 2009

J Clin Microbiol

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To improve the clinical outcome of Staphylococcus aureus septicemia, the early selection of appropriate antibiotic treatment is crucial. Molecular diagnostics represents an attractive approach for the rapid identification of S. aureus and the determination of its methicillin (meticillin) resistance. In direct comparison to other molecular assays (sa442 and mecA real-time PCRs) and standard laboratory procedures, we evaluated the BD GeneOhm StaphSR assay for its use in the detection of S. aureus and methicillin-resistant S. aureus (MRSA) from spiked blood culture bottles (n ‫؍‬ 134). In the case of detecting S. aureus (n ‫؍‬ 90; for methicillin-susceptible S. aureus, n ‫؍‬ 45; for MRSA, n ‫؍‬ 45), the BD GeneOhm StaphSR assay had a sensitivity and a specificity of 100% each (95% confidence intervals [CIs], 96.0 to 100% and 82.4 to 100%, respectively). For MRSA (n ‫؍‬ 45), the test was 95.6% (95% CI, 84.9 to 99.5%) sensitive and 95.3% (95% CI, 86.9 to 99.0%) specific. Overall, five discrepant results arose with this assay due to the presence of methicillin-susceptible, revertant MRSA strains (3/45) and MRSA strains that were not detected by the BD GeneOhm StaphSR assay (2/45). Compared to other real-time PCR-based molecular approaches and to conventional standard laboratory methods, the BD GeneOhm StaphSR turned out to be an appropriate diagnostic tool for a rapid (ϳ1.5 h), preliminary identification of S. aureus and MRSA from blood cultures.

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Accuracy and Potential Usefulness of Triplex Real-Time PCR for Improving Antibiotic Treatment of Patients with Blood Cultures Showing Clustered Gram-Positive Cocci on Direct Smears

Cited by 56 publications

References 45 publications

Intestinal carriage of Extended Spectrum Beta-Lactamase producing E. coli in women with urinary tract infections, Cameroon

Intestinal carriage of Extended Spectrum Beta-Lactamase producing E. coli in women with urinary tract infections, Cameroon

Rapid Detection of bla _KPC Carbapenemase Genes by Internally Controlled Real-Time PCR Assay Using Bactec Blood Culture Bottles

Evaluation of the BD GeneOhm StaphSR Assay for Detection of Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Isolates from Spiked Positive Blood Culture Bottles

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Accuracy and Potential Usefulness of Triplex Real-Time PCR for Improving Antibiotic Treatment of Patients with Blood Cultures Showing Clustered Gram-Positive Cocci on Direct Smears

Cited by 56 publications

References 45 publications

Intestinal carriage of Extended Spectrum Beta-Lactamase producing E. coli in women with urinary tract infections, Cameroon

Intestinal carriage of Extended Spectrum Beta-Lactamase producing E. coli in women with urinary tract infections, Cameroon

Rapid Detection of bla KPC Carbapenemase Genes by Internally Controlled Real-Time PCR Assay Using Bactec Blood Culture Bottles

Evaluation of the BD GeneOhm StaphSR Assay for Detection of Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Isolates from Spiked Positive Blood Culture Bottles

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Rapid Detection of bla _KPC Carbapenemase Genes by Internally Controlled Real-Time PCR Assay Using Bactec Blood Culture Bottles