Phenotypic Detection of Carbapenemase Production in Carbapenem-Resistant Enterobacteriaceae by Modified Hodge Test and Modified Strip Carba NP Test

Abstract

Objective Carbapenems are last resort antibiotics for multidrug-resistant Enterobacteriaceae. However, resistance to carbapenem is increasing at an alarming rate worldwide leading to major therapeutic failures and increased mortality rate. Early and effective detection of carbapenemase producing carbapenem-resistant Enterobacteriaceae (CRE) is therefore key to control dissemination of carbapenem resistance in nosocomial as well as community-acquired infection. The aim of present study was to evaluate efficacy of Modified strip Carba NP (CNP) test against Modified Hodge test (MHT) for early detection of carbapenemase producing Enterobacteriaceae (CPE).

Material and Methods Enterobacteriaceae isolated from various clinical samples were screened for carbapenem resistance. A total of 107 CRE were subjected to MHT and Modified strip CNP test for the detection of CPE.

Statistical Analysis It was done on Statistical Package for the Social Sciences (SPSS) software, IBM India; version V26. Nonparametric test chi-square and Z-test were used to analyze the results within a 95% level of confidence.

Results Out of 107 CRE, 94 (88%) were phenotypically confirmed as carbapenemase producer by Modified strip CNP test and 46 (43%) were confirmed by Modified Hodge Test (MHT). Thirty-eight (36%) isolates showed carbapenemase production by both MHT and CNP test, 56 isolates (52%) were CNP test positive but MHT negative, eight (7%) isolates were MHT positive but CNP test negative and five (5%) isolates were both MHT and CNP test negative. There is statistically significant difference in efficiency of Modified CNP test and MHT (p < 0.05).

Conclusion Modified strip CNP test is simple and inexpensive test which is easy to perform and interpret and gives rapid results in less than 5 minutes. It has high degree of sensitivity and specificity. Modified strip CNP test shows significantly higher detection capacity for carbapenemase producers as compared with MHT.

Keywords

carbapenem-resistant Enterobacteriaceae - carbapenemase producing Enterobacteriaceae - modified Hodge test - modified strip Carba NP test

• References

• 1 Koneman EW. Koneman’s color atlas and textbook of Diagnostic Microbiology, Characteristics for Presumptive Identification of Enterobacteriaceae. 7th ed. Philadelphia, PA:Wolters Kluwer;2017:685–689
• 2 Blair JMA, Webber MA, Baylay AJ, Ogbolu DO, Piddock LJV. Molecular mechanisms of antibiotic resistance. Nat Rev Microbiol 2015; 13 (01) 42-51
• 3 Peter-Getzlaff S, Polsfuss S, Poledica M. et al. Detection of AmpC beta-lactamase in Escherichia coli: comparison of three phenotypic confirmation assays and genetic analysis. J Clin Microbiol 2011; 49 (08) 2924-2932
• 4 Papp-Wallace KM, Endimiani A, Taracila MA, Bonomo RA. Carbapenems: past, present, and future. Antimicrob Agents Chemother 2011; 55 (11) 4943-4960
• 5 Yigit H, Queenan AM, Anderson GJ. et al. Novel carbapenem-hydrolyzingβ-lactamase, KPC-1, from a carbapenem-resistant strain of Klebsiella pneumoniae . Antimicrob Agents Chemother 2001; 45 (04) 1151-1161
• 6 World Health Organization. Guidelines for the prevention and control of carbapenem-resistant Enterobacteriaceae, Acinetobacter baumannii and Pseudomonas aeruginosa in health care facilities. 2017:ISBN 978–92–4-155017–8. Available at: https://www.who.int/infection-prevention/publications/guidelines-cre/en/. Accessed November 17, 2019
• 7 Clinical and Laboratory Standards Institute. AST news update. 2017; 1 (02) 9
• 8 Tille PM. Bailey & Scotts Diagnostic Microbiology, Expanded Spectrum Cephalosporin Resistance & Carbapenemase Resistance. 13th ed. St. Louis: Elsevier. 2014: 326
• 9 Cornaglia G, Giamarellou H, Rossolini GM. Metallo-β-lactamases: a last frontier forβ-lactams?. Lancet Infect Dis 2011; 11 (05) 381-393
• 10 Nordmann P, Poirel L, Walsh TR, Livermore DM. The emerging NDM carbapenemases. Trends Microbiol 2011; 19 (12) 588-595
• 11 Poirel L, Potron A, Nordmann P. OXA-48-like carbapenemases: the phantom menace. J Antimicrob Chemother 2012; 67 (07) 1597-1606
• 12 van D Duin, Doi Y. The global epidemiology of carbapenemase-producing Enterobacteriaceae. Virulence 2017; 8 (04) 460-469
• 13 Kazi M, Drego L, Nikam C. et al. Molecular characterization of carbapenem-resistant Enterobacteriaceae at a tertiary care laboratory in Mumbai. Eur J Clin Microbiol Infect Dis 2015; 34 (03) 467-472
• 14 Shanmugam P, Meenakshisundaram J, Jayaraman P. blaKPC gene detection in clinical isolates of carbapenem resistant Enterobacteriaceae in a tertiary care hospital. J Clin Diagn Res 2013; 7 (12) 2736-2738
• 15 Nordmann P, Poirel L, Dortet L. Rapid detection of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis 2012; 18 (09) 1503-1507
• 16 Poirel L, Walsh TR, Cuvillier V, Nordmann P. Multiplex PCR for detection of acquired carbapenemase genes. Diagn Microbiol Infect Dis 2011; 70 (01) 119-123
• 17 Cuzon G, Naas T, Bogaerts P, Glupczynski Y, Nordmann P. Evaluation of a DNA microarray for the rapid detection of extended-spectrumβ-lactamases (TEM, SHV and CTX-M), plasmid-mediated cephalosporinases (CMY-2-like, DHA, FOX, ACC-1, ACT/MIR and CMY-1-like/MOX) and carbapenemases (KPC, OXA-48, VIM, IMP and NDM).. J Antimicrob Chemother 2012; 67 (08) 1865-1869
• 18 Hrabák J, Walková R, Studentová V, Chudácková E, Bergerová T. Carbapenemase activity detection by matrix-assisted laser desorption ionization-time of flight mass spectrometry. J Clin Microbiol 2011; 49 (09) 3222-3227
• 19 Lee W, Chung HS, Lee Y. et al. Comparison of matrix-assisted laser desorption ionization-time-of-flight mass spectrometry assay with conventional methods for detection of IMP-6, VIM-2, NDM-1, SIM-1, KPC-1, OXA-23, and OXA-51 carbapenemase-producing Acinetobacter spp., Pseudomonas aeruginosa, and Klebsiella pneumoniae. Diagn Microbiol Infect Dis 2013; 77 (03) 227-230
• 20 Poirel L, Nordmann P. Rapidec Carba NP Test for rapid detection of carbapenemase producers. J Clin Microbiol 2015; 53 (09) 3003-3008
• 21 Girlich D, Poirel L, Nordmann P. Value of the modified Hodge test for detection of emerging carbapenemases in Enterobacteriaceae. J Clin Microbiol 2012; 50 (02) 477-479
• 22 Carvalhaes CG, Picão RC, Nicoletti AG, Xavier DE, Gales AC. Cloverleaf test (modified Hodge test) for detecting carbapenemase production inKlebsiella pneumoniae: be aware of false positive results. J Antimicrob Chemother 2010; 65 (02) 249-251
• 23 Anderson KF, Lonsway DR, Rasheed JK. et al. Evaluation of methods to identify the Klebsiella pneumoniae carbapenemase in Enterobacteriaceae. J Clin Microbiol 2007; 45 (08) 2723-2725
• 24 Mochon AB, Garner OB, Hindler JA. et al. New Delhi metallo-β-lactamase (NDM-1)-producing Klebsiella pneumoniae: case report and laboratory detection strategies. J Clin Microbiol 2011; 49 (04) 1667-1670
• 25 Srisrattakarn A, Lulitanond A, Wilailuckana C. et al. Modification and evaluation of the Carba NP test by use of paper strip for simple and rapid detection of carbapenemase-producing Enterobacteriaceae. World J Microbiol Biotechnol 2016; 32 (07) 117
• 26 Ho PL, Wang Y, Wing-Sze Tse C. et al. Rapid detection of carbapenemase production in Enterobacteriaceae by use of a modified paper strip Carba NP method. J Clin Microbiol 2017; 56 (01) e01110-e01117
• 27 Centersfor Disease Control and Prevention. Modified Hodge test for carbapenemase detection in Enterobacteriaceae. Available at: https://www.cdc.gov/HAI/pdfs/labSettings/HodgeTest_Carbapenemase_Enterobacteriaceae.pdf. Accessed November 17, 2019
• 28 Bedenić B, Sardelić S. Textbook on Carbapenemases. London:Intech Open;2018:1–12
• 29 Nair PK, Vaz MS. Prevalence of carbapenem resistant Enterobacteriaceae from a tertiary care hospital in Mumbai, India. J Microbiol Inf Dis 2013; 3 (04) 207-210
• 30 Chauhan K, Pandey A, Asthana AK, Madan M. Evaluation of phenotypic tests for detection of Klebsiella pneumoniae carbapenemase and metallo-beta-lactamase in clinical isolates of Escherichia coli and Klebsiella species. Indian J Pathol Microbiol 2015; 58 (01) 31-35
• 31 Pandurangan S, BegumEsak S, Narayanasamy A. et al. Phenotypic detection methods of carbapenemase production in Enterobacteriaceae. Int J Curr Microbiol Appl Sci 2015; 4 (06) 547-552
• 32 Mangayarkarasi V, Moses SP, Swarna SR. et al. In-house standardization of Carba NP test for carbapenemase detection in gram negative bacteria. Int J Curr Microbiol Appl Sci 2018; 7 (01) 2876-2881
• 33 Datta S, Dey R, Dey JB, Ghosh S. A comparative study of modified Hodge test and Carba NP test for detecting carbapenemase production in gram-negative bacteria. Med J Dr D.Y. Patil Uni. 2017; 10: 365-369
• 34 Bartolini A, Frasson I, Cavallaro A, Richter SN, Palù G. Comparison of phenotypic methods for the detection of carbapenem non-susceptible Enterobacteriaceae . Gut Pathog 2014; 6: 13
• 35 Centers for Disease Control and Prevention. Case definition for surveillance of CP-CRE; 2018. Available at: https://wwwn.cdc.gov/nndss/conditions/carbapenemase-producing-carbapenem-resistant-enterobacteriaceae/case-definition/2018/. Accessed November 17, 2019
• 36 Vasoo S, Cunningham SA, Kohner PC. et al. Comparison of a novel, rapid chromogenic biochemical assay, the Carba NP test, with the modified Hodge test for detection of carbapenemase-producing Gram-negative bacilli. J Clin Microbiol 2013; 51 (09) 3097-3101
• 37 Yamada K, Kashiwa M, Arai K, Nagano N, Saito R. Comparison of the Modified-Hodge test, Carba NP test, and carbapenem inactivation method as screening methods for carbapenemase-producingEnterobacteriaceae . J Microbiol Methods 2016; 128: 48-51
• 38 Giske CG, Martinez-Martinez L, Cantón R, Stefani S, Skov R, Glupczynski Y, et al. European Committee on Antimicrobial Susceptibility Testing. Guidelines for detection of resistance mechanisms and specific resistances of clinical and/or epidemiological importance. 2017;2:1–43
• 39 Tijet N, Boyd D, Patel SN, Mulvey MR, Melano RG. Evaluation of the Carba NP test for rapid detection of carbapenemase-producing Enterobacteriaceae and Pseudomonas aeruginosa. Antimicrob Agents Chemother 2013; 57 (09) 4578-4580