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采用更新后的临床和实验室标准协会(Clinical and Laboratory Standards Institute)折点,通过纸片扩散法、VITEK 2以及肉汤微量稀释法,比较环丙沙星和氨基糖苷类药物对头孢曲松不敏感肠杆菌科细菌的药敏试验。

Comparison of ciprofloxacin and aminoglycoside susceptibility testing for ceftriaxone non-susceptible Enterobacterales by disk diffusion and VITEK 2 vs. broth microdilution using updated Clinical and Laboratory Standards Institute breakpoints.

作者信息

Sohani Zahra N, Lieu Anthony, Semret Makeda, Cheng Matthew P, Simic Nancy, Bamba Reggie, Patel Mina, Lawandi Alexander, Lee Todd C

机构信息

Division of Infectious Diseases, Department of Medicine, McGill University Health Centre, Montréal, QC, Canada.

Division of Medical Microbiology, Department of Laboratory Medicine, McGill University Health Centre, Montréal, Canada.

出版信息

BMC Microbiol. 2025 Mar 31;25(1):175. doi: 10.1186/s12866-025-03923-7.

Abstract

BACKGROUND

Fluoroquinolones and aminoglycosides are potential treatment choices in the setting of increasingly multi-drug resistant Enterobacterales. The Clinical & Laboratory Standards Institute (CLSI) breakpoints for fluoroquinolones and aminoglycosides in the Enterobacterales were revised in 2019 and 2022, respectively. However, performance of existing widely used automated systems, such as the VITEK 2 AST-N391 card, has not been extensively tested for MDR isolates at these new breakpoints.

OBJECTIVE

To assess performance of the new breakpoints for ciprofloxacin, gentamicin, and tobramycin on the VITEK 2 system (bioMérieux, France) and disk diffusion by comparing to broth microdilution for ceftriaxone nonsusceptible Enterobacterales.

METHODS

Ninety-four ceftriaxone non-susceptible Escherichia coli and Klebsiella pneumoniae isolates were identified between January 2021-June 2023. Broth microdilution was used as the reference standard against which disk diffusion and VITEK 2 susceptibility testing were compared. For the Vitek 2, we used the AST-N391 card and interpreted the results according to the updated CLSI breakpoints.

RESULTS

Overall, 22.3% of isolates were susceptible to ciprofloxacin by BMD. Compared to BMD, disk diffusion had an overall minor error rate of 7.4% (95%CI 3.0-14.7%) with 0 major or very major errors (97.5% CI 0-3.8%). For the VITEK 2, a minor error rate of 13.8% (95% CI 7.6-22.5%), major error rate 19.0% (95%CI 7.7-40.0%) and very major error rate 0% (97.5%CI 0-3.8%) was noted. By comparison, 69.1% and 56.4% of isolates were susceptible to gentamicin and tobramycin, respectively. Disk diffusion and the VITEK 2 system both correctly categorized 100% of gentamicin susceptible and non-susceptible isolates. For tobramycin, disk diffusion had a 3.2% rate of misclassification (all minor errors) and the VITEK 2 had 2.1% rate of misclassification (all minor errors). There were no major or very major errors.

CONCLUSIONS

Our findings suggest that both disk diffusion and to a greater extent the AST-N391 card for the VITEK 2 system will overcall non-susceptibility according to current CLSI breakpoints for ciprofloxacin. By contrast, the existing AST-N391 VITEK 2 card can likely be used to correctly infer susceptibility to gentamicin and tobramycin.

摘要

背景

在对多种药物耐药的肠杆菌科细菌日益增多的情况下,氟喹诺酮类和氨基糖苷类药物是潜在的治疗选择。临床和实验室标准协会(CLSI)分别于2019年和2022年修订了肠杆菌科细菌中氟喹诺酮类和氨基糖苷类药物的折点。然而,现有的广泛使用的自动化系统,如VITEK 2 AST-N391卡片,在这些新折点下对多重耐药菌株的性能尚未得到广泛测试。

目的

通过将头孢曲松不敏感的肠杆菌科细菌的纸片扩散法和VITEK 2系统与肉汤微量稀释法进行比较,评估环丙沙星、庆大霉素和妥布霉素新折点在VITEK 2系统(法国生物梅里埃公司)上的性能。

方法

在2021年1月至2023年6月期间鉴定出94株头孢曲松不敏感的大肠埃希菌和肺炎克雷伯菌菌株。以肉汤微量稀释法作为参考标准,将纸片扩散法和VITEK 2药敏试验与之进行比较。对于Vitek 2,我们使用AST-N391卡片,并根据更新后的CLSI折点解释结果。

结果

总体而言,通过肉汤微量稀释法,22.3%的菌株对环丙沙星敏感。与肉汤微量稀释法相比,纸片扩散法的总体小错误率为7.4%(95%CI 3.0-14.7%),无主要或非常主要错误(97.5%CI 0-3.8%)。对于VITEK 2系统,小错误率为13.8%(95%CI 7.6-22.5%),主要错误率为19.0%(95%CI 7.7-40.0%),非常主要错误率为0%(97.5%CI 0-3.8%)。相比之下,69.1%和56.4%的菌株分别对庆大霉素和妥布霉素敏感。纸片扩散法和VITEK 2系统均正确分类了100%的庆大霉素敏感和不敏感菌株。对于妥布霉素,纸片扩散法的错误分类率为3.2%(均为小错误),VITEK 2系统的错误分类率为2.1%(均为小错误)。无主要或非常主要错误。

结论

我们的研究结果表明,根据当前CLSI环丙沙星折点,纸片扩散法以及在更大程度上VITEK 2系统的AST-N391卡片会过度判定为不敏感。相比之下,现有的VITEK 2 AST-N391卡片可能可用于正确推断对庆大霉素和妥布霉素的敏感性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9f71/11956328/20afab75f70d/12866_2025_3923_Fig1_HTML.jpg

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