2009, Number 4
Rapid detection of ESBL-producing gram-negative bacteria isolated from blood: a reasonable and reliable tool for middle and low resource countries
PDF size: 341.96 Kb.
ABSTRACTIntroduction. Delay in appropriate treatment in patients with bacteraemia can increase morbidity, mortality, and health expenditures. We compared the Rapid Direct Test (RDT) designed to detect ESBL producing gram negative bac teria (GNB) directly from positive blood cultures bottles, with two conventional ESBL detection tests: Screening and Confir matory Disk Diffusion Assay (SC DDA) and an MIC Screen ing and ESBL E test (MIC/ET). Material and methods. We screened all blood cultures in a tertiary care facility from August to December 2005. We only included one positive bot tle per patient in which GNB were observed. RDT: Blood from each bottle was inoculated on Mueller Hinton agar. Ceftazi dime and cefotaxime disks with and without clavulanic acid were added and incubated at 35 ºC ± 2 ºC for 24 h. Results were interpreted according to CLSI recommendations for the SC DDA and MIC/ET. All methods were performed simulta neously. Time for reporting as an ESBL producer and cost of the tests were recorded. Results. We isolated 124 GNB in 114 episodes of bacteraemia, 10 of them (8.8%) polymicrobial; 79 (63.7%) of the GNB were enteric bacteria, 44 (35.5%) glucose non fermenter GNB and one Haemophilus influenzae. The most common microorganism was Escherichia coli in 56 epi sodes (45.2%), followed by Pseudomonas aeruginosa in 24 (19.3%), and Klebsiella pneumoniae in 13 (10.5%). Of the 114 episodes, 41 (36%) had at least one GNB resistant to 3rd gen eration cephalosporins, and 25 (21.9%) were caused by an ESBL producing GNB. Sensitivity, specificity, positive predic tive value (PPV) and negative predictive value (NPV) for the RDT were 96%, 98.9%, 96% and 98.9%, respectively. Agree ment by kappa index between RDT and SC DDA was 0.95 and between the RDT and MIC/ET was 0.92. The RDT detected 24 25 ESBL producing bacteria. The mean time to detect an iso late as an ESBL producer after a positive blood culture bottle signal was 1.02 ± 0.19 days when using the RDT, and 3.40 ± 0.59 days when using any other method. The difference in re porting time was 2.38 ± 0.63 days (p ‹ 0.0001). Our estimat ed cost per test was $1.54 for RDT, $2.32 for screening/ confirmatory SC DDA, and $49.65 for MIC screening and MIC/ET. Conclusions. The RDT is a rapid, reliable and easy analysis to perform, as well as cost effective.
Kliebe C, Nies BA, Meyer JF, Tolxdorff Neutzling RM, Wiede mann B. Evolution of plasmid coded resistance to broad spectrum cephalosporins. Antimicrob Agents Chemother 1985; 28: 302 7.
Bradford PA. Extended spectrum β lactamases in the 21st cen tury: characterization, epidemiology, and detection of this im portant resistance threat. Clin Microbiol Rev 2001; 14: 933 51.
Poirel L, Gniadkowski M, Nordmann P. Biochemical analysis of the ceftazidime hydrolyzing extended spectrum b lactamase CTX M 15 and of its structurally related β lactamase CTX M 3. J Antimicrob Chemother 2002; 50: 1031 4.
Turner PJ. Extended spectrum B lactamases. Clin Infect Dis 2005; 41(Suppl. 4): S273 S275.
Biedenbach DJ, Moet GJ, Jones RN. Occurrence and antimi crobial resistance pattern comparisons among bloodstream in fection isolates from the SENTRY Antimicrobial Surveillance Program (1997 2002). Diag Microbiol Infect Dis 2004; 50: 59 69.
Turner PJ, Greenhalgh JM, Edwards JR, McKellar J. The MYS TIC (Meropenem Yearly Susceptibility Test Information Collec tion) programme. Int J Antimicrob Agents 1999; 13: 117 25.
Silva J, Gatica R, Aguilar C, Becerra Z, Garza Ramos U, Veláz quez M, et al. Outbreak of infection with extended spectrum β lactamase producing Klebsiella pneumoniae in a Mexican Hospital. J Clin Microbiol 2001; 39: 3193 6.
Alcántar D, Tinoco JC, Gayosso C, Carlos A, Daza C, Pérez Orado MC, et al. Nosocomial bacteraemia and urinary tract in fections caused by extended spectrum β lactamase producing Klebsiella pneumoniae with plasmids carrying both SHV 5 and TLA 1 genes. Clin Infect Dis 2004; 38: 1067 74.
Mosqueda Gómez JL, Montaño A, Rolon AL, Cervantes C, Bobadilla del Valle M, Silva J, et al. Molecular epidemiology and risk factors of bloodstream infections caused by extended spectrum β lactamase producer Klebsiella pneumoniae. A case control study. Int J Infect Dis 2008; 12: 653 9.
Clinical and Laboratory Standards Institute/NCCLS (2008). Performance standards for antimicrobial susceptibility testing; 18th Informational Supplement. CLSI/NCCLSI document M100 S18, Vol. 27, No. 1 [ISBN 1 56238 556 9] Clinical and Laboratory Standards Institute, 940 West Valley Road, Suite 1400, Wayne, Pennsylvania 19087 1898, USA.
Du B, Long Y, Liu H, Chen D, Liu D, Xu Y, et al. Extended spectrum beta lactamase producing Escherichia coli and Kleb siella pneumoniae bloodstream infection: risk factors and clinical outcome. Intensive Care Med 2002; 28: 1718 23.
Ibrahim EH, Sherman G, Ward S, Fraser VJ, Kollef MH. The Influence of inadequate antimicrobial treatment of bloodstream infections on patient outcomes in the ICU setting. Chest 2000; 118: 146 55.
Navon Venezia S, Ben Ami R, Schwaber MJ, Leavitt A, Schwartz D, Carmeli Y. Protocol for the accelerated detection of extended spectrum beta lactamase producing Escherichia coli and Klebsiella pneumoniae strains from blood cultures. Eur J Clin Microbiol Infect Dis 2004; 23: 200 02.
Navon Venezia S, Leavitt A, Ben Ami R, Aharoni Y, Schwaber MJ, Schwartz D, et al. Evaluation of an accelerated protocol for detection of extended spectrum beta lactamase producing gram negative bacilli from positive blood cultures. J Clin Microbiol 2005; 43: 439 41.
Swenson JM, Hindler JF, Jorgensen JH. Special phenotypic methods for detecting antibacterial resistance. In: Murray PR, Baron EJ, Jorgensen JH, Pfaller MA, Yolken RH (eds.). Ma nual of clinical microbiology. 8th Ed. Washington D.C., US: ASM Press; 2003, p. 1178 95.
Ozakin C, Sinirtas M, Sevgican E, Kazak E, Gedikoglu S. Comparison of the E test method with an automated bacterial identification and antimicrobial susceptibility detection system for screening extended spectrum beta lactamase producers. Scand J Infect Dis 2003; 35: 700 03.
Pfaller MA, Segreti J. Overview of the epidemiological profile and laboratory detection of extended spectrum β lactamases. Clin Infect Dis 2006; 42(Suppl. 4): S153 S163.
Vercauteren E, Descheemaeker P, Ieven M, Sanders CC, Goos sens H. Comparison of screening methods for detection of ex tended spectrum b lactamases and their prevalence among blood isolates of Escherichia coli and Klebsiella spp. in a Bel gian teaching hospital. J Clin Microbiol 1997; 35: 2191 7.
Spanu T, Sanguinetti M, Tumbarello M, D’Inzeo T, Posteraro B, Santangelo R, et al. Evaluation of the new Vitek 2 exten ded spectrum beta lactamases (ESBL) test for rapid detection of ESBL production in Enterobacteriaceae isolates. J Clin Microbiol 2006; 44: 3257 62.
Lautenbach E, Patel JB, Bilker WB, Edelstein PH, Fishman NO. Extended spectrum beta lactamase producing Escheri chia coli and Klebsiella pneumoniae: risk factors for infec tion and impact of resistance on outcomes. Clin Infect Dis 2001; 32: 1162 71.
Rodríguez Baño J, Navarro MD, Romero L, Muniain MA, Pe rea EJ, Perez Cano R, et al. Clinical and molecular epidemiolo gy of extended spectrum B lactamase producing Escherichia coli as a cause of nosocomial infection or colonization: impli cations for control. Clin Infect Dis 2006; 42: 37 45.
Schwaber MJ, Navon Venezia S, Kaye KS, Ben Ami R, Schwartz D, Carmeli Y. Clinical and economic impact of bac teraemia with extended spectrum beta lactamase producing Enterobacteriaceae. Antimicrob Agents Chemother 2006; 50: 1257 62.
González Vértiz A, Alcántar Curiel D, Cuauhtli M, Daza C, Gayosso C, Solache G, et al. Multiresistant extended spectrum β lactamase producing Klebsiella pneumoniae causing an out break of nosocomial bloodstream infection. Infect Control Hosp Epidemiol 2001; 22: 725 8.
Ndugulile F, Jureen R, Harthung S, Urassa W, Langeland N. Extended spectrum β lactamases among gram negative bacteria of nosocomial origin from an intensive care unit of a tertiary health facility in Tanzania. BMC Infect Dis 2005; 5: 86.
Pokharel BM, Koirala J, Dahal RK, Mishra SK, Khadga PK, Tuladhar NR. Multidrug resistant and extended spectrum beta lactamase (ESBL) producing Salmonella enterica (serotypes Typhi and Paratyphi A) from blood isolates in Nepal: survei llance of resistance and a search for newer alternatives. Int J Infect Dis 2006; 10: 434 8.
Kato Maeda M, Bautista Alavez A, Rolón Montes de Oca AL, Ramos Hinojosa A, Ponce de León A, Bobadilla del Valle M, et al. Increasing trend of antimicrobial drug resistance in orga nisms causing bacteremia at a tertiary care hospital: 1995 to 2000. Rev Invest Clin 2003; 55: 600 05.