Biyokimyasal Testler
Özet
Karbapenem grubu antibiyotiklere dirençli bakterilerin neden olduğu enfeksiyonlar, hem halk sağlığı hem de ekonomik açıdan ciddi tehditler oluşturmaktadır. Karbapenemlere direnç mekanizmaları arasında en önemlisi karbapenemaz enzimlerinin üretimidir. Bu enzimler, karbapenem antibiyotikleri etkisiz hale getirerek tedavi seçeneklerini sınırlar. Karbapenemaz enzimlerinin saptanması için moleküler ve fenotipik yöntemler kullanılmakta olup, fenotipik yöntemler arasında biyokimyasal (kolorimetrik) testler öne çıkmaktadır. Bu testlerin çalışma ilkesi, karbapenem grubu bir antibiyotik olan imipenemin ß-laktam halkasının karbapenemazın enzimatik aktivitesi ile hidrolizi sonucu pH indikatörünün renk değişimine dayanmaktadır. En yaygın biyokimyasal test olan Carba NP test hızlı, ucuz ve uygulanması kolay olmakla birlikte OXA-48 ve GES tipi karbapenemazlar gibi bazı enzim gruplarında düşük duyarlılığa sahiptir. Bu sınırlamaları gidermek için Blue-Carba, Rapidec Carba NP, β-CARBA, MAST Carba PAcE, CarbAcineto NP gibi çeşitli türevleri ve ticari kitler geliştirilmiştir. Bu testlerin duyarlılık ve özgüllük oranları kullanılan bakteri türüne, karbapenemaz tipine ve test koşullarına göre değişiklik göstermektedir. Bazı testler, karbapenemaz sınıflarını ayırt edebilme yeteneğine sahipken, çoğu sadece varlığı saptayabilmektedir. Bu nedenle test sonuçları dikkatle değerlendirilmelidir. Moleküler doğrulama testleriyle desteklenmesi, tanı doğruluğunu artıracaktır. Sonuç olarak, biyokimyasal karbapenemaz testleri, özellikle kaynak kısıtlı laboratuvarlarda uygun maliyetli ve pratik tanı araçlarıdır. Bununla birlikte, testlerin sınırlamaları göz önünde bulundurulmalı ve gerektiğinde tamamlayıcı yöntemlerle desteklenmelidir.
Infections caused by microorganisms resistant to carbapenem antibiotics pose serious threats to both public health and the economy. Among the mechanisms of resistance to carbapenems, the most important is the production of carbapenemase enzymes. These enzymes render carbapenem antibiotics ineffective, thereby limiting treatment options. Molecular and phenotypic methods are used to detect carbapenemase enzymes, with biochemical (colorimetric) tests being the most prominent among phenotypic methods. The principle of these tests is based on the color change of a pH indicator resulting from the hydrolysis of the β-lactam ring of imipenem, a carbapenem antibiotic, by the enzymatic activity of carbapenemase. The most common biochemical test, the Carba NP test, is fast, inexpensive, and easy to perform, but it has low sensitivity for certain enzyme groups, such as OXA-48 and GES-type carbapenemases. To overcome these limitations, various modifications and commercial kits have been developed, such as Blue-Carba, Rapidec Carba NP, β-CARBA, MAST Carba PAcE, and CarbAcineto NP. The sensitivity and specificity rates of these tests vary depending on the type of bacteria, carbapenemase type, and test conditions. While some tests can distinguish between carbapenemase classes, most can only detect their presence. Therefore, test results should be carefully evaluated. Support from molecular confirmation tests will increase diagnostic accuracy. In conclusion, biochemical carbapenemase tests are cost-effective and practical diagnostic tools, especially in resource-limited laboratories. However, the limitations of the tests should be considered, and they should be supported by complementary methods when necessary.
Referanslar
Zhong H, Lu Wu M, Feng WJ, Huang SF, Yang P. Accuracy and applicability of different phenotypic methods for carbapenemase detection in Enterobacteriaceae: A systematic review and meta-analysis J Glob Antimicrob Resist 2020; 21:138-147.
Suriya R V, KV L, Feliciana J H, Aishwarya R. Diagnostic Test Precision of Modified Carbapenem Inactivation Method and Carbapenemase Nordmann-Poirel Test for Phenotypic Detection of Carbapenemase Production in Enterobacterales: A Systematic Review. Cureus 2024; 6(8): e67322.
Tamma PD, Opene BNA, Gluck A, Chambers KK, Carroll KC, Simner PJ. Comparison of 11 phenotypic assays for accurate detection of carbapenemase producing Enterobacteriaceae. J Clin Microbiol 2017; 55:1046-1055.
Bouslah Z. Carba NP test for the detection of carbapenemase-producing Pseudomonas aeruginosa. Med Mal Infect 2020;50(6):466-479.
Lifshitz Z, Adler A, Carmeli Y. Comparative study of a novel biochemical assay, the Rapidec Carba NP test, for detecting carbapenemase producing Enterobacteriaceae. J Clin Microbiol 2016; 54:453–456.
European Committee on Antimicrobial Susceptibility Testing (EUCAST): Klinik ve/veya epidemiyolojik önemi olan direnç mekanizmaları ve direnç özelliklerini saptama kılavuzu Versiyon 2.0. 2017 (www.tmc-online.org/userfiles/file/EUCAST-ceviri-2017.pdf).
Nordmann P, Poirel L, Dortet L. Rapid detection of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis 2012;18(9):1503–7.
Rao MR, Chandrashaker P, Mahale RP, Shivappa SG, Gowda RS, Chitharagi VB. Detection of carbapenemase production in Enterobacteriaceae and Pseudomonas species by carbapenemase Nordmann–Poirel test. J Lab Physicians 2019;11:107-10.
Tamma PD, Simner PJ. Phenotypic detection of carbapenemase producing organisms from clinical isolates. J Clin Microbiol 2018; 56:e01140-18.
Literacka E, Herda M, Baraniak A, et al. Evaluation of the Carba NP test for carbapenemase detection in Enterobacte-riaceae, Pseudomonas spp. and Acinetobacter spp., and its practical use in theroutine work of a national reference laboratory for susceptibility testing. Eur J Clin Microbiol Infect Dis 2017;36(11):2281–7.
CLSI Performance Standards for Antimicrobial Susceptibility Testing. In:Wayne PA, editor. CLSI supplement M100S. 25th edition Pennsylvania, USA:Clinical and Laboratory Standards Institute; 2015.
Pires J, Novais A, Peixe L. Blue-carba, an easy biochemical test for detection of diverse carbapenemase producers directly from bacterial cultures. J Clin Microbiol 2013;51(12):4281-3.
Bernabeu S, Dortet L, Naas T. Evaluation of the b-CARBATM test, a colorimetric test for the rapid detection of carbapenemase activity in Gram-negative bacilli. J Antimicrob Chemother 2017; 72: 1646–1658.
Rezzoug I, Emeraud C, Sauvadet A, Cotellon G, Naas T, Dortet L. Evaluation of the MAST PAcE colorimetric test for rapid detection of carbapenemase activity in Gram negative bacilli. Antimicrob Agents Chemother 2021; 65:e02351-20.
Simner PJ, Johnson JK, Brasso WB, et al. Multicenter evaluation of the modified carbapenem inactivation method and the Carba NP for detection of carbapenemase-producing Pseudomonas aeruginosa and Acinetobacter baumannii. J Clin Microbiol 2018; 56:e1369-17.
Gniadek TJ, Carroll KC, Simner PJ. Carbapenem resistant non-glucose fermenting Gram-negative bacilli: the missing piece to the puzzle. J Clin Microbiol 2016; 54:1700–1710.
Dortet L, Poirel L, Errera C, Nordmann P. CarbAcineto NP Test for Rapid Detection of Carbapenemase-Producing Acinetobacter spp. J Clin Microbiol 2014;52(7):2359-64
Dortet L, Poirel L, Nordmann P. Rapid identification of carbapenemase types inEnterobacteriaceae and Pseudomonas spp. by using a biochemical test. Antimi-crob Agents Chemother 2012;56(12):6437-40.
Referanslar
Zhong H, Lu Wu M, Feng WJ, Huang SF, Yang P. Accuracy and applicability of different phenotypic methods for carbapenemase detection in Enterobacteriaceae: A systematic review and meta-analysis J Glob Antimicrob Resist 2020; 21:138-147.
Suriya R V, KV L, Feliciana J H, Aishwarya R. Diagnostic Test Precision of Modified Carbapenem Inactivation Method and Carbapenemase Nordmann-Poirel Test for Phenotypic Detection of Carbapenemase Production in Enterobacterales: A Systematic Review. Cureus 2024; 6(8): e67322.
Tamma PD, Opene BNA, Gluck A, Chambers KK, Carroll KC, Simner PJ. Comparison of 11 phenotypic assays for accurate detection of carbapenemase producing Enterobacteriaceae. J Clin Microbiol 2017; 55:1046-1055.
Bouslah Z. Carba NP test for the detection of carbapenemase-producing Pseudomonas aeruginosa. Med Mal Infect 2020;50(6):466-479.
Lifshitz Z, Adler A, Carmeli Y. Comparative study of a novel biochemical assay, the Rapidec Carba NP test, for detecting carbapenemase producing Enterobacteriaceae. J Clin Microbiol 2016; 54:453–456.
European Committee on Antimicrobial Susceptibility Testing (EUCAST): Klinik ve/veya epidemiyolojik önemi olan direnç mekanizmaları ve direnç özelliklerini saptama kılavuzu Versiyon 2.0. 2017 (www.tmc-online.org/userfiles/file/EUCAST-ceviri-2017.pdf).
Nordmann P, Poirel L, Dortet L. Rapid detection of carbapenemase-producing Enterobacteriaceae. Emerg Infect Dis 2012;18(9):1503–7.
Rao MR, Chandrashaker P, Mahale RP, Shivappa SG, Gowda RS, Chitharagi VB. Detection of carbapenemase production in Enterobacteriaceae and Pseudomonas species by carbapenemase Nordmann–Poirel test. J Lab Physicians 2019;11:107-10.
Tamma PD, Simner PJ. Phenotypic detection of carbapenemase producing organisms from clinical isolates. J Clin Microbiol 2018; 56:e01140-18.
Literacka E, Herda M, Baraniak A, et al. Evaluation of the Carba NP test for carbapenemase detection in Enterobacte-riaceae, Pseudomonas spp. and Acinetobacter spp., and its practical use in theroutine work of a national reference laboratory for susceptibility testing. Eur J Clin Microbiol Infect Dis 2017;36(11):2281–7.
CLSI Performance Standards for Antimicrobial Susceptibility Testing. In:Wayne PA, editor. CLSI supplement M100S. 25th edition Pennsylvania, USA:Clinical and Laboratory Standards Institute; 2015.
Pires J, Novais A, Peixe L. Blue-carba, an easy biochemical test for detection of diverse carbapenemase producers directly from bacterial cultures. J Clin Microbiol 2013;51(12):4281-3.
Bernabeu S, Dortet L, Naas T. Evaluation of the b-CARBATM test, a colorimetric test for the rapid detection of carbapenemase activity in Gram-negative bacilli. J Antimicrob Chemother 2017; 72: 1646–1658.
Rezzoug I, Emeraud C, Sauvadet A, Cotellon G, Naas T, Dortet L. Evaluation of the MAST PAcE colorimetric test for rapid detection of carbapenemase activity in Gram negative bacilli. Antimicrob Agents Chemother 2021; 65:e02351-20.
Simner PJ, Johnson JK, Brasso WB, et al. Multicenter evaluation of the modified carbapenem inactivation method and the Carba NP for detection of carbapenemase-producing Pseudomonas aeruginosa and Acinetobacter baumannii. J Clin Microbiol 2018; 56:e1369-17.
Gniadek TJ, Carroll KC, Simner PJ. Carbapenem resistant non-glucose fermenting Gram-negative bacilli: the missing piece to the puzzle. J Clin Microbiol 2016; 54:1700–1710.
Dortet L, Poirel L, Errera C, Nordmann P. CarbAcineto NP Test for Rapid Detection of Carbapenemase-Producing Acinetobacter spp. J Clin Microbiol 2014;52(7):2359-64
Dortet L, Poirel L, Nordmann P. Rapid identification of carbapenemase types inEnterobacteriaceae and Pseudomonas spp. by using a biochemical test. Antimi-crob Agents Chemother 2012;56(12):6437-40.
