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.
Leal HF, Azevedo J, Silva GE, Amorim AM, de Roma LR, Arraes AC, et al. Bloodstream infections caused by multidrug-resistant gram-negative bacteria: epidemiological, clinical and microbiological features. BMC Infect Dis 2019; 19(1): 1–1.
Santoro A, Franceschini E, Meschiari M, Menozzi M, Zona S, Venturelli C, et al. Epidemiology and risk factors associated with mortality in consecutive patients with bacterial bloodstream infection: impact of MDR and XDR bacteria. Open Forum Infect Dis 2020; 7: ofaa461.
Sands K, Carvalho MJ, Portal E, Thomson K, Dyer C, Akpulu C. Characterization of antimicrobial-resistant gram-negative bacteria that cause neonatal sepsis in seven low-and middle-income countries. Nat Microbiol 2021; 6(4): 512–23.
Hassuna NA, AbdelAziz RA, Zakaria A, Abdelhakeem M. Extensively-drug resistant Klebsiella pneumoniae recovered from neonatal sepsis cases from a major NICU in Egypt. Front Microbiol 2020; 11: 1375.
Ghaith DM, Zafer MM, Said HM, Elanwary S, Elsaban S, Al-Agamy MH, et al. Genetic diversity of carbapenem-resistant Klebsiella pneumoniae causing neonatal sepsis in intensive care unit, Cairo, Egypt. Eur J Clin Microbiol Infect Dis 2020; 39: 583–591.
Lee YL, Chen HM, Hii M, Hsueh PR. Carbapenemase-producing Enterobacterales infections: recent advances in diagnosis and treatment. Int J Antimicrob Agents 2022; 59(2): 106528.
Logan LK, Weinstein RA. The epidemiology of carbapenem-resistant Enterobacteriaceae: the impact and evolution of a global menace. J Infect Dis 2017; 215(suppl_1): S28–36.
Tang Y, Xu C, Xiao H, Wang L, Cheng Q, Li X. Gram-negative bacteria bloodstream infections in patients with hematological malignancies–the impact of pathogen type and patterns of antibiotic resistance: a retrospective cohort study. Infect Drug Resist 2021; 12: 3115–24.
Baer D, Azrad M, Saleh N, Peretz A. Detection of carbapenem-resistant enterobacterales in simulated blood culture in 15 minutes. Life 2021; 11(2): 145.
Banerjee R, Humphries R. Clinical and laboratory considerations for the rapid detection of carbapenem-resistant Enterobacteriaceae. Virulence 2017; 8(4): 427–39.
Boutal H, Vogel A, Bernabeu S, Devilliers K, Creton E, Cotellon G, et al. A multiplex lateral flow immunoassay for the rapid identification of NDM-, KPC-, IMP-and VIM-type and OXA-48-like carbapenemase-producing Enterobacteriaceae. J Antimicrob Chemother 2018; 3(4): 909–15.
Hong J, Kang D, Kim D. Performance evaluation of the newly developed in vitro rapid diagnostic test for detecting OXA-48-like, KPC-, NDM-, VIM-and IMP-type carbapenemases: the RESIST-5 OKNVI multiplex lateral flow assay. Antibiotics 2021; 10(4): 460.
Kumar M, Tandel K, Shergill SP, Bhalla GS, Mahajan P, Swarnim V, et al. Rapid detection of carbapenem resistance among gram-negative organisms directly from positive blood culture bottles. Med J Armed Forces India 2023; 79(3): 267–74.
Taha R, Kader O, Shawky S, Rezk S. Ceftazidime-Avibactam plus aztreonam synergistic combination tested against carbapenem-resistant Enterobacterales characterized phenotypically and genotypically: a glimmer of hope. Ann Clin Microbiol Antimicrob 2023; 122(1): 21.
Abbasi E, Ghaznavi-Rad E. High frequency of NDM-1 and OXA-48 carbapenemase genes among Klebsiella pneumoniae isolates in central Iran. BMC Microbiol 2023; 23(1): 98.
El-Kholy AA, Girgis SA, Shetta MA, Abdel-Hamid DH, Elmanakhly AR. Molecular characterization of multidrug-resistant gram-negative pathogens in three tertiary hospitals in Cairo, Egypt. Eur J Clin Microbiol Infect Dis 2020; 39: 987–92.
Kriger O, Shatzman-Steuerman R, Smollan G, Belausov N, Sarkisian G, Amit S. Rapid detection of carbapenemase-producing enterobacteriaceae from blood culture bottles of known CPE carriers: real-world experience. J Pediatr Infect Dis 2022 Jan 1; 41(1): 45–7.
Hamprecht A, Vehreschild JJ, Seifert H, Saleh A. Rapid detection of NDM, KPC and OXA-48 carbapenemases directly from positive blood cultures using a new multiplex immunochromatographic assay. PLoS One 2018; 13(9): e0204157.
Lima-Morales DD, Ávila H, Soldi T, Dalmolin TV, Lutz L, Aquino V, et al. Rapid detection of carbapenemase production directly from blood culture by colorimetric methods: evaluation in a routine microbiology laboratory. J Clin Microbiol 2018; 56(9): 10–128.
Meier M, Hamprecht A. Systematic comparison of four methods for detection of carbapenemase-producing Enterobacterales directly from blood cultures. J Clin Microbiol 2019; 57(11): 10–128.
Weinstein MP, Clinical and Laboratory Standards Institute. Performance standards for antimicrobial susceptibility testing; 2021.
Meheissen M, Okasha H. Verification of direct methods for detection of carbapenemase producing Enterobacteriaceae from blood cultures. Microbes Infect Dis 2022; 3(3): 687–692.
De Cueto M, Ceballos E, Martinez-Martinez L, Perea EJ, Pascual A. Use of positive blood cultures for direct identification and susceptibility testing with the Vitek 2 system. J Clin Microbiol 2004; 42(8): 3734–8.
Micó M, Navarro F, de Miniac D, González Y, Brell A, López C, et al. Efficacy of the FilmArray blood culture identification panel for direct molecular diagnosis of infectious diseases from samples other than blood. J Med Microbiol 2015; 64(12): 1481–8.
Ramsamy Y, Mlisana KP, Amoako DG, Abia AL, Ismail A, Allam M, et al. Mobile genetic elements-mediated Enterobacterales-associated carbapenemase antibiotic resistance genes propagation between the environment and humans: a One Health South African study. Sci Total Environ 2022; 1(806): 150641.