Diagnostic accuracy of resazurin based rapid test for Colistin susceptibility in Carbapenem resistant Acinetobacter baumanii isolated form ICU settings

Sidrah  Muqarrab; Irfan Ali  Mirza; Sakeenah Hussain  Naqvi; Adeel Gardezi; Muhammad  Roshan; Rabia  Sajjad; Anam  Imtiaz

doi:10.51253/pafmj.v75iSUPPL-2.10171

Authors

Sidrah Muqarrab Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi/National University of Medical Sciences (NUMS) Pakistan
Irfan Ali Mirza Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi/National University of Medical Sciences (NUMS) Pakistan
Sakeenah Hussain Naqvi Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi/National University of Medical Sciences (NUMS) Pakistan
Adeel Gardezi Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi/National University of Medical Sciences (NUMS) Pakistan
Muhammad Roshan Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi/National University of Medical Sciences (NUMS) Pakistan
Rabia Sajjad Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi/National University of Medical Sciences (NUMS) Pakistan
Anam Imtiaz Department of Microbiology, Armed Forces Institute of Pathology Rawalpindi/National University of Medical Sciences (NUMS) Pakistan

DOI:

https://doi.org/10.51253/pafmj.v75iSUPPL-2.10171

Keywords:

Acinetobacter Baumannii, Colistin Susceptibility, Rapid Resazurin Test

Abstract

Objective: To determine diagnostic accuracy of rapid resazurin based test for Colistin susceptibility testing in Carbapenem resistant Acinetobacter baumannii isolated from ICU patients.

Study Design: Validation study.

Place and Duration of Study: Department of Microbiology, Armed Forces Institute of Pathology, Rawalpindi Pakistan, from Jul to Dec 2022.

Methodology: A resazurin based rapid test for Colistin susceptibility testing in Acinetobacter baumannii was assessed using a total of 100 clinical isolates from ICU patients. This test detects the presence of viable bacterial cells in Mueller-Hinton broth containing a predefined concentration of Colistin sulfate. Bacterial suspensions of the isolates are added to Colistin containing Mueller-Hinton broth on a 96-well polystyrene microtiter plate and incubated for 3 hours at 35° ± 2°C. Resazurin (a viability colorant) is then added to the wells and results are interpreted on basis of color change (from blue to pink in resistant isolates) after a total of 4 hours.

Results: The test accurately detected resistance in all 12 resistant isolates and had a sensitivity and specificity of 100% and 97.7% respectively. False positive results were seen in 2 out of 88 sensitive isolates. The overall diagnostic accuracy of the test was 98%.

Conclusion: This resazurin based test is in-expensive, easy to perform, accurate and provides earlier results in just 4 hours as compared to 16-24 hours with the routine lab methods. This test can serve as a good alternate to conventional methods in resource limited settings and for infection control purposes.

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References

Vázquez-López R, Solano-Gálvez SG, Juárez Vignon-Whaley JJ, Abello Vaamonde JA, Padró Alonzo LA, Rivera Reséndiz A, et al. Acinetobacter baumannii Resistance: A Real Challenge for Clinicians. Antibiotics 2020; 9(4).

Kyriakidis I, Vasileiou E, Pana ZD, Tragiannidis A. Acinetobacter baumannii Antibiotic Resistance Mechanisms. Pathogens 2021; 10(3).

Ramirez MS, Bonomo RA, Tolmasky ME. Carbapenemases: Transforming Acinetobacter baumannii into a Yet More Dangerous Menace. Biomolecules 2020; 10(5).

WHO publishes list of bacteria for which new antibiotics are urgently needed [Internet]. [cited 2023 Mar 17]. Available from:

https://www.who.int/news/item/27-02-2017-who-publishes-list-of-bacteria-for-which-new-antibiotics-are-urgently-needed

Nguyen M, Joshi SG. Carbapenem resistance in Acinetobacter baumannii, and their importance in hospital‐acquired infections: a scientific review. J Appl Microbiol 2021; 131(6): 2715–2738.

Tacconelli E, Carrara E, Savoldi A, Harbarth S, Mendelson M, Monnet DL, et al. Discovery, research, and development of new antibiotics: the WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect Dis 2018; 18(3): 318–327.

Hamidian M, Nigro SJ. Emergence, molecular mechanisms and global spread of carbapenem-resistant Acinetobacter baumannii. Microb Genomics [Internet]. 2019 Oct 1 [cited 2023 Mar 20];5(10). Available from:

https://www.microbiologyresearch.org/content/journal/mgen/10.1099/mgen.0.000306

Mohd Sazlly Lim S, Zainal Abidin A, Liew SM, Roberts JA, Sime FB. The global prevalence of multidrug-resistance among Acinetobacter baumannii causing hospital-acquired and ventilator-associated pneumonia and its associated mortality: A systematic review and meta-analysis. J Infect 2019; 79(6): 593–600.

Khalid F, Saleem S, Ahmad I. High prevalence of carbapenem resistant Acinetobacter baumannii associated respiratory tract infections in Pakistani hospitals. J Pak Med Assoc 2020; (0): 1.

Ahsan U, Mushtaq F, Saleem S, Malik A, Sarfaraz H, Shahzad M, et al. Emergence of high colistin resistance in carbapenem resistant Acinetobacter baumannii in Pakistan and its potential management through immunomodulatory effect of an extract from Saussurea lappa. Front Pharmacol 2022; 13: 986802.

Piperaki ET, Tzouvelekis LS, Miriagou V, Daikos GL. Carbapenem-resistant Acinetobacter baumannii: in pursuit of an effective treatment. Clin Microbiol Infect 2019; 25(8): 951–957.

Isler Burcu, Doi Yohei, Bonomo Robert A., Paterson David L. New Treatment Options against Carbapenem-Resistant Acinetobacter baumannii Infections. Antimicrob Agents Chemother 2018; 63(1): e01110-18.

Jung H, Pitout JDD, Mitton BC, Strydom KA, Kingsburgh C, Coetzee J, et al. Evaluation of the rapid ResaPolymyxin Acinetobacter/Pseudomonas NP test for rapid colistin resistance detection in lactose non-fermenting Gram-negative bacteria [Internet]. Vol. 70, Journal of Medical Microbiology. Microbiology Society; 2021. Available from:

https://www.microbiologyresearch.org/content/journal/jmm/10.1099/jmm.0.001373

Turlej-Rogacka A, Xavier BB, Janssens L, Lammens C, Zarkotou O, Pournaras S, et al. Evaluation of colistin stability in agar and comparison of four methods for MIC testing of colistin. Eur J Clin Microbiol Infect Dis 2018; 37: 345–353.

EUCAST. Recommendations for MIC determination of colistin (polymyxin E) as recommended by the joint CLSI-EUCAST Polymyxin Breakpoints Working Group [Internet]. EUCAST; 2016 Mar [cited 2023 Mar 23]. Available from:

http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/General_documents/Recommendations_for_MIC_determination_of_colistin_March_2016.pdf

Lescat Mathilde, Poirel Laurent, Tinguely Camille, Nordmann Patrice. A Resazurin Reduction-Based Assay for Rapid Detection of Polymyxin Resistance in Acinetobacter baumannii and Pseudomonas aeruginosa. J Clin Microbiol 2019; 57(3): e01563-18.

O’brien J, Wilson I, Orton T, Pognan F. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur J Biochem 2000; 267(17): 5421–5426.

Bouvier M, Sadek M, Pomponio S, D’Emidio F, Poirel L, Nordmann P. RapidResa Polymyxin Acinetobacter NP® Test for Rapid Detection of Polymyxin Resistance in Acinetobacter baumannii. Antibiotics 2021; 10(5).

Tahseen U, Talib MT. Acinetobacter infections as an emerging threat in intensive care units. J Ayub Med Coll Abbottabad 2015; 27(1): 113–116.

CLSI. Performance Standards for Antimicrobial Susceptibility Testing. 32rd ed. Wayne, PA: Clinical and Laboratory Standards Institute; 2022. M100-Ed32.

Nordmann P, Jayol A, Poirel L. Rapid Detection of Polymyxin Resistance in Enterobacteriaceae. Emerg Infect Dis 2016; 22(6): 1038–1043.

Jia H, Fang R, Lin J, Tian X, Zhao Y, Chen L, et al. Evaluation of resazurin-based assay for rapid detection of polymyxin-resistant gram-negative bacteria. BMC Microbiol 2020; 20(1): 7.

Kon H, Abramov S, Amar Ben Dalak M, Elmaliach N, Schwartz D, Carmeli Y, et al. Performance of Rapid PolymyxinTM NP and Rapid PolymyxinTM Acinetobacter for the detection of polymyxin resistance in carbapenem-resistant Acinetobacter baumannii and Enterobacterales. J Antimicrob Chemother. 2020; 75(6): 1484–1490.

Germ J, Poirel L, Kisek TC, Spik VC, Seme K, Premru MM, et al. Evaluation of resazurin-based rapid test to detect colistin resistance in Acinetobacter baumannii isolates. Eur J Clin Microbiol Infect Dis 2019; 38(11): 2159–2162