The Diagnostic Capacity of Three Phenotypic Techniques of Extended-Spectrum β-Lactamase Detection

Abera Abdeta, A. Bitew, Surafel Fentaw, Estifanos Tsige, D. Assefa, E. Tigabu, T. Lejisa, Yordanos Kefyalew, Ebisa Fekede
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Abstract

Background: Early detection of extended-spectrum β-lactamases (ESBLs) producing bacteria is critical for infection prevention and control. Numerous phenotypic approaches and automated systems have been developed for detecting ESBL bacteria. However, there is a scarcity of data in Ethiopia regarding the most reliable, simple, and cost-effective methods for detecting ESBL-producing bacteria. This study, therefore, aimed to evaluate the diagnostic performance of three phenotypic approaches for detecting ESBL-producing bacteria. Methods: In this study, 117 isolates of Klebsiella pneumoniae, Escherichia coli, Klebsiella oxytoca, and Proteus mirabilis were examined. Cefotaxime (30 µg) and ceftazidime (30 µg) were used for screening ESBL enzymes. A screening breakpoints of≤27 mm and≤22 mm were used for cefotaxime (30 µg) and ceftazidime (30 µg), respectively, as per the Clinical and Laboratory Standards Institute (CLSI) guidelines. All 117 strains were further confirmed by the Vitek 2 compact, double disk synergy, ESBL Epsilometer test, and combined disk method. The combined disk method was adopted as the reference method. Results: Out of 117 isolates, 90 (86%) had zone diameters of≤27 mm and≤22 mm for cefotaxime (30 µg) and ceftazidime (30 µg), respectively. The reference method detected 76 (65%) ESBL isolates out of 117 ones. From among the three techniques (i.e., double disk synergy, Vitek 2 compact, and ESBL Epsilometer test), the double disk synergy method demonstrated overall sensitivity and specificity of 97.4% and 97.6%, respectively. Vitek-2, cefotaxime, and ceftazidime Epsilometer test indicated indeterminate results of 6.8%, 6.8%, and 5.1% respectively. Conclusion: Double disk synergy was found to have the highest sensitivity and specificity for detecting ESBL isolates with no indeterminate results.
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三种增谱β-内酰胺酶检测表型技术的诊断能力
背景:早期发现产生广谱β-内酰胺酶(ESBLs)的细菌是预防和控制感染的关键。许多表型方法和自动化系统已经开发用于检测ESBL细菌。然而,埃塞俄比亚缺乏关于检测产生esbl的细菌的最可靠、最简单和最具成本效益的方法的数据。因此,本研究旨在评估检测产esbl细菌的三种表型方法的诊断性能。方法:对117株肺炎克雷伯菌、大肠埃希菌、产氧克雷伯菌和奇异变形杆菌进行检测。用头孢噻肟(30µg)和头孢他啶(30µg)筛选ESBL酶。根据临床和实验室标准协会(CLSI)指南,头孢噻肟(30µg)和头孢他啶(30µg)的筛选断点分别为≤27 mm和≤22 mm。采用Vitek 2紧凑法、双圆盘协同法、ESBL Epsilometer试验和联合圆盘法对117株进行进一步鉴定。采用组合圆盘法作为参考方法。结果:117株分离菌中,头孢噻肟(30µg)和头孢噻啶(30µg)的区径分别为≤27 mm和≤22 mm,占86%;参考方法在117株ESBL中检出76株(65%)。在双盘协同法、Vitek 2 compact法和ESBL Epsilometer法三种检测方法中,双盘协同法的总灵敏度和特异度分别为97.4%和97.6%。Vitek-2、头孢噻肟和头孢他啶Epsilometer试验的不确定结果分别为6.8%、6.8%和5.1%。结论:双盘协同法检测ESBL分离株具有最高的灵敏度和特异性,无不确定结果。
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