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Rapid detection of carbapenemase-producing gram-negative bacteria directly from blood culture bottles for a reliable guided empiric therapy

Shahinda Rezk, Nourhan Ashraf, Hassan Heshmat, Gamal Elsawaf, Amel Elsheredy
Acta Microbiol Immunol Hung . 2024 Apr 4;71(2):140-147

Abstract
Bloodstream infections (BSIs) caused by multidrug-resistant bacteria are a critical life-threatening challenge which necessitates the urgency to trigger life-saving treatment in a timely manner. This study aimed to evaluate the time required for rapid detection of carbapenemase-producing Enterobacterales (CPE) directly from blood culture bottles to optimize empirical treatment of BSI, especially in pediatric and infant patients, using a cost-effective method. This study included 419 Gram-negative bacteria, of which Klebsiella pneumoniae and Escherichia coli were the most common CPE causing BSI in pediatric and neonatal patients. Phenotypic and genotypic resistance of the selected isolates (45 K. pneumoniae and 9 E. coli) were determined by VITEK-2 Compact system and PCR, respectively. BACT/ALERT bottles were spiked with isolates. Finally, colorimetric RESIST-BC assay and Vitek-2 compact system were evaluated for the rapid detection of carbapenem-resistant bacteria directly from positive blood culture bottles. All selected isolates were phenotypically resistant to carbapenems. PCR showed that blaNDM and blaOXA-48 were present in all isolates, blaVIM was present in 44.4%, while blaKPC and blaIMP were entirely absent. The RESIST-BC kit showed good agreement with PCR for blaNDM and blaOXA-48, demonstrating high sensitivity and specificity, but not with blaVIM. These findings point out that RESIST-BC assay demonstrated an exceptionally short detection time for CPE, completing all cases within the first hour after the blood culture bottles flagged positive. It is also superior in providing a clue for clinicians on antibiotic combinations that can be administered, depending on the type of β-lactamases detected, promptly and efficiently, with low expenses.

https://pubmed.ncbi.nlm.nih.gov/38573768/

Application and clinical impact of the RESIST-4 O.K.N.V. rapid diagnostic test for carbapenemase detection in blood cultures and clinical samples

Roth S, Berger FK, Link A, Nimmesgern A, Lepper PM, Murawski N, Bittenbring JT, Becker SL.
Eur J Clin Microbiol Infect Dis. 2021 Feb;40(2):423-428

Abstract

Invasive infections caused by carbapenemase-producing bacteria are associated with excess mortality. We applied a rapid diagnostic test (RDT) on clinical samples with an elevated likelihood of carbapenemase-producing bacteria and documented its impact on antibiotic treatment decisions. Among 38 patients, twelve tested positive for infections caused by carbapenemase-producing bacteria (31.6%), mainly in blood cultures. KPC (n = 10) was more frequent than OXA-48 (n = 2). RDT-based carbapenemase detection led to a treatment modification to ceftazidime/avibactam-containing regimens in all patients before detailed antibiotic testing results became available. Eleven patients (92%) survived the acute infection, whereas one patient with a ceftazidime/avibactam- and colistin-resistant OXA-48-positive isolate died.

Direct detection of carbapenem resistance determinants in clinical specimens using immunochromatographic lateral flow devices.

Wareham DW, Phee LM, Abdul Momin MHF.
J Antimicrob Chemother. 2018 Jul 1;73(7):1997-1998

No abstract available

Rapid detection of NDM, KPC and OXA-48 carbapenemases directly from positive blood cultures using a new multiplex immunochromatographic assay.

Hamprecht A, Vehreschild JJ, Seifert H, Saleh A.
PLoS One. 2018 Sep 14;13(9):e0204157.

Abstract

Bloodstream infections caused by carbapenemase-producing Enterobacteriaceae (CPE) are associated with treatment failure and increased mortality. Detection of CPE from blood cultures (BC) by standard methods takes 16-72 hours, which can delay the initiation of appropriate antimicrobial therapy and compromise patient outcome. In the present study, we developed and evaluated a new method for the rapid detection of carbapenemases directly from positive BC using a new multiplex immunochromatographic test (ICT). The new ICT was assessed using 170 molecularly characterized Enterobacteriaceae clinical isolates including 126 CPE (OXA-48-like (N = 79), KPC (N = 18) and NDM (N = 29)). After spiking with bacteria and incubation in a BC system, blood from positive BC bottles was hemolyzed, bacteria concentrated by centrifugation and lysed. The lysate was transferred to the RESIST-3 O.K.N. ICT (Coris BioConcept, Gembloux, Belgium), which detects OXA-48-like, KPC and NDM carbapenemases. The final results of the ICT were read when they became positive, at the latest after 15 min. All CPE isolates (126/126) were correctly detected with the new protocol (100% sensitivity, 100% specificity). There was perfect concordance between ICT results and molecular characterization. Total time to result was 20-45 min.

P0810- Rapid detection of carbapenemase-producing Enterobactyeriaceae directly from positive blood cultures by a new immunochromatographic assay

A. Hamprecht, H. Seifert and A. Saleh
28th European Congress of Clinical Microbiology and Infectious Diseases, April 21 - 24 April 2018
Poster

P2325- Rapid detection of carbapenemase-producing organisms directly from positive blood cultures using a combined MALDI-TOF MS and RESIST O.K.N. workflow

E. Wey, L. Ainsworth, T. McHugh and I. Balakrishnan
28th European Congress of Clinical Microbiology and Infectious Diseases, April 21 - 24 April 2018
Poster

Detection of OXA-370 directly from rectal swabs and blood culture vials using an immunochromatographic assay.

Nodari CS, Gales AC, Barth AL, Magagnin CM, Zavascki AP, Carvalhaes CG.
J Microbiol Methods. 2017 Aug;139:92-94

Abstract

We evaluated the performance of OXA-48 K-SeT assay for detecting OXA-370 directly from spiked rectal swabs and blood culture vials. The limit of detection of this test was 104UFC/mL for rectal swabs. Detection of the OXA-370-producing isolates was successfully achieved directly from positive blood culture vials independent of growing conditions.

The prevalence of Extended-Spectrum B-lactamse (ESBL) and carbapenem-resistant Enterobacteriaceae (CRE) isolates in positive blood cultures of patients in a teaching hospital in Kuwait over a 2-year period.

Taqi M, Jamal W and Rotimi V
Open Forum Infectious Diseases, Volume 4, Issue suppl_1, 2017, S1944

Abstract

Invasive infections caused by carbapenemase-producing bacteria are associated with excess mortality. We applied a rapid diagnostic test (RDT) on clinical samples with an elevated likelihood of carbapenemase-producing bacteria and documented its impact on antibiotic treatment decisions. Among 38 patients, twelve tested positive for infections caused by carbapenemase-producing bacteria (31.6%), mainly in blood cultures. KPC (n = 10) was more frequent than OXA-48 (n = 2). RDT-based carbapenemase detection led to a treatment modification to ceftazidime/avibactam-containing regimens in all patients before detailed antibiotic testing results became available. Eleven patients (92%) survived the acute infection, whereas one patient with a ceftazidime/avibactam- and colistin-resistant OXA-48-positive isolate died.
https://academic.oup.com › ofid › S564 › ofx163.1476.pdf