Rapid Detection of Carbapenemase Production Directly from Blood Culture by Colorimetric Methods: Evaluation in a Routine Microbiology Laboratory

Lima-Morales, Daiana de; Ávila, Helena; Soldi, Tatiana; Dalmolin, Tanise Vendruscolo; Lutz, Larissa; Aquino, Valério Rodrigues; Zavascki, Alexandre Prehn; Barth, Afonso Luís

doi:10.1128/jcm.00325-18

Cited by 21 publications

(15 citation statements)

References 16 publications

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“…A range of strategies may be useful in achieving this goal. According to the study results, our hospitals implemented the following recommendations for ICU patients: (i) regular rectal screening [36,37] will be useful to improve this clinical strategy. We found no differences in 30-day mortality rates among patients who received colistin-base, CAZ-AVI-based, or other treatment regimens.…”

Section: Discussionmentioning

confidence: 99%

Time to appropriate antibiotic therapy is a predictor of outcome in patients with bloodstream infection caused by KPC-producing Klebsiella pneumoniae

et al. 2020

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Background: Bloodstream infections (BSIs) by Klebsiella pneumoniae carbapenemase (KPC)-producing Klebsiella pneumoniae (Kp) are associated with high mortality. The aim of this study is to assess the relationship between time to administration of appropriate antibiotic therapy and the outcome of patients with BSI due to KPC-Kp hospitalized in intensive care unit (ICU). Methods: An observational study was conducted in the ICUs of two academic centers in Italy. Patients with KPC-Kp bacteremia hospitalized between January 2015 to December 2018 were included. The primary outcome was the relationship between time from blood cultures (BC) collection to appropriate antibiotic therapy and 30-day mortality. The secondary outcome was to evaluate the association of different treatment regimens with 30-day mortality and a composite endpoint (30-day mortality or nephrotoxicity). A Cox regression analysis to identify factors independently associated with 30-day mortality was performed. Hazard ratio (HR) and 95% confidence interval (CI) were calculated. Results: A total of 102 patients with KPC-Kp BSI were included. The most common sources of infection were intraabdominal (23.5%), urinary tract (20.6%), and skin and skin structure (17.6%). The 30-day mortality was 45%. Median time to appropriate antibiotic therapy was shorter in patients who survived (8.5 h [IQR 1-36]) versus those who died (48 h [IQR 5-108], p = 0.014). A propensity score matching showed that receipt of an in vitro active therapy within 24 h from BC collection was associated with lower 30-day mortality (HR = 0.36, 95% CI: 0.188-0.690, p = 0.0021). At Cox regression analysis, factors associated with 30-day mortality were primary bacteremia (HR 2.662 [95% CI 1.118-6.336], p = 0.027), cardiovascular disease , p = 0.029), time (24-h increments) from BC collection to appropriate therapy (HR 1.382 [95% CI 1.132-1.687], p = 0.001), SOFA score (HR 1.122 [95% CI 1.036-1.216], p = 0.005), and age (HR 1.030 [95% CI 1.006-1.054], p = 0.012). Ceftazidime-avibactam-containing regimens were associated with reduced risk of composite endpoint (30-day mortality OR nephrotoxicity) (HR 0.231 [95% CI 0.071-0.745], p = 0.014) compared to colistin-containing regimens.Conclusions: Time to appropriate antibiotic therapy is an independent predictor of 30-day mortality in patients with KPC-Kp BSI. Appropriate antibiotic therapy should begin within 24 h from the collection of BC.

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Section: Discussionmentioning

confidence: 99%

Time to appropriate antibiotic therapy is a predictor of outcome in patients with bloodstream infection caused by KPC-producing Klebsiella pneumoniae

et al. 2020

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“…3 Double-disk synergy test for detection of strains belongs to class A β-lactamases, Modified Hodge test to detect class A β-lactamases and meropenem E-test strip to identify class B β-lactamases is common tests. 13,14 One of the most critical advantages of phenotypic methods is their cost-effectiveness. Of course, the fatality of these methods and the low speed of diagnosis should be mentioned from its main disadvantages, and the need to use sensitive and rapid molecular methods along with these phenotypic methods is essential.…”

Section: Introductionmentioning

confidence: 99%

<p>Distribution of Class B and Class A β-Lactamases in Clinical Strains of <em>Pseudomonas aeruginosa</em>: Comparison of Phenotypic Methods and High-Resolution Melting Analysis (HRMA) Assay</p>

Dehbashi¹,

Tahmasebi²,

Alikhani³

et al. 2020

IDR

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Background: There are various phenotypic methods for identifying class B and class A βlactamase enzymes in Pseudomonas aeruginosa. The purpose of this study was to compare the sensitivity and specificity of different phenotypic methods with HRMA assay to detect βlactamase-producing P. aeruginosa strains. Methods: Eighty-eight of P. aeruginosa isolates were collected from different specimens. Conventional double-disk test (DDT) and EDTA-imipenem microbiological (EIM) were performed to detect ESBL and MBL-producing strains, respectively. Meanwhile, the Modified Hodge test and Carba-NP test were performed on all carbapenem-resistant strains. HRMA method and sensitivity and specificity of primers were determined based on the melt curve temperature range. In all comparisons, PCR was considered as the gold standard. Results: Of the 402 isolates collected from different clinical specimens, 88 isolates of P. aeruginosa were identified. However, 43 strains were (48.88%) ESBL-producing, and 7 strains (7.95%) were MBL-producing. Also, using the Modified Hodge test and Carba-NP method, 11 (12.5%) and 19 (21.59%) strains were carbapenemase-producing, respectively. The results of the HRMA test revealed that genes coding for bla SHV , bla TEM , bla KPC , bla IMP , bla VIM, and bla GES were detected in 44.31%, 22.72%, 13.63%, 14.7%, 5.6%, and 2.27% of P. aeruginosa isolates. Nonetheless, for bla KPC and bla GES genes, sensitivity and specificity of the Carba-NP test were 90.47%, 94.87%, and 83.36%, 94.80%, respectively. However, sensitivity and specificity of MHT was 91.66%, 98.70%, and 77.77%, 96.42%, respectively. For bla SHV and bla TEM genes, sensitivity and specificity of DDT were 95.55%, 95.55%, and 86%, 83.50%, respectively. However, sensitivity and specificity of EMI were 77.77%, 97.59%, and 91.66%, 97.43% for bla VIM and bla IMP , respectively. Conclusion: The HRMA is a powerful, accurate, closed-tube, rapid method for detecting β-lactamase genes in P. aeruginosa. The high sensitivity and specificity of this method, along with phenotypic tests, play a useful role in increasing the predictive value of clinical reports.

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“…An interesting finding was the decreased diagnostic sensitivity for the detection of VIM activity in anaerobic versus aerobic bottles (53.6% and 100%, respectively), raising the hypothesis that this enzyme may be downregulated, inactivated, or rapidly degraded in anaerobic bottles. Previous studies have shown high sensitivity values when metallo-␤-lactamases are detected directly from BCs by colorimetric methods (8)(9)(10)(11), immunochromatographic tests (14,16), or MALDI-TOF-based hydrolysis assays (17,18), even though these screening were performed only on spiked aerobic BC bottles. More recently, Cointe et al have reported the low reproducibility of the RESIST-4 O.K.N.V.…”

Section: Resultsmentioning

confidence: 99%

“…Although a number of genotypic diagnostic methods for the detection of ␤-lactamase-encoding genes in blood cultures (BCs) are currently available, they are expensive procedures requiring skilled personnel and are not able to identify all ESBL-or carbapenemase-encoding genes (6). Thus, during the last decade, several phenotype-based assays have been developed to detect ESBL or carbapenemase production in BCs, including colorimetric methods (7)(8)(9)(10)(11)(12)(13), immunochromatogenic assays (14)(15)(16), and ␤-lactam hydrolysis assays combined with matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) (17)(18)(19)(20).…”

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confidence: 99%

Direct β-Lactam Inactivation Method: a New Low-Cost Assay for Rapid Detection of Carbapenemase- or Extended-Spectrum-β-Lactamase-Producing Enterobacterales Directly from Positive Blood Culture Bottles

et al. 2019

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We validate and evaluate a new phenotypic assay, named the direct β-lactam inactivation method (dBLIM), for the rapid and simultaneous detection of carbapenemase or extended-spectrum-cephalosporinase activity directly from Enterobacterales (EB)-positive blood cultures (BCs). It originates from the carbapenem inactivation method (CIM), an inexpensive and highly sensitive assay for carbapenemase activity detection. dBLIM cutoff values to detect extended-spectrum β-lactamase (ESBL) and carbapenemase activities resulted in diameters of ≤12 mm for a 5-μg-cefotaxime disk and for a 10-μg-meropenem disk. dBLIM assessment was determined with both aerobic and anaerobic BC bottles spiked with 422 characterized EB strains, classifiable into the following 4 phenotypic groups: (i) ESBL/AmpC-type β-lactamase (ACBL)/carbapenemase (CARB)-nonproducing (np-ESBL/ACBL/CARB) EB (n = 116), (ii) ESBL-producing EB (n = 111), (iii) AmpC-β-lactamase-producing EB (n = 33), and (iv) carbapenemase-producing EB (n = 162). No false-positive results were obtained in any of the np-ESBL/ACBL/CARB EB, ESBL, and AmpC groups, demonstrating an overall assay specificity of 100%. There were no significant discrepancies in dBLIM performance between aerobic and anaerobic BCs across all groups, except with VIM-type carbapenemase-expressing EB. Interestingly, among BCs spiked with blaVIM-harboring EB, the sensitivity rates of the assay in anaerobic and aerobic bottles were 53.6% and 100%, respectively. In contrast, dBLIM performance was deemed excellent for the KPC, OXA-48, and NDM carbapenemase producers regardless of the type of bottle being tested, with a sensitivity rate ranging between 99% and 100%. Concerning the detection of the extended-spectrum cephalosporinases of the ESBL-producing and AmpC types, dBLIM sensitivities was 100% and 84 to 87%, respectively. dBLIM may be a cost-effective and highly robust phenotypic screening method for the reliable detection of carbapenemases or extended-spectrum cephalosporinases directly from BCs on the same day of bottle positivity detection.

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Rapid Detection of Carbapenemase Production Directly from Blood Culture by Colorimetric Methods: Evaluation in a Routine Microbiology Laboratory

Cited by 21 publications

References 16 publications

Time to appropriate antibiotic therapy is a predictor of outcome in patients with bloodstream infection caused by KPC-producing Klebsiella pneumoniae

Time to appropriate antibiotic therapy is a predictor of outcome in patients with bloodstream infection caused by KPC-producing Klebsiella pneumoniae

<p>Distribution of Class B and Class A β-Lactamases in Clinical Strains of <em>Pseudomonas aeruginosa</em>: Comparison of Phenotypic Methods and High-Resolution Melting Analysis (HRMA) Assay</p>

Direct β-Lactam Inactivation Method: a New Low-Cost Assay for Rapid Detection of Carbapenemase- or Extended-Spectrum-β-Lactamase-Producing Enterobacterales Directly from Positive Blood Culture Bottles

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