MALDI-TOF MS直接靶板微滴生长测定法在CRKP快速检测中的应用

doi:10.3969/j.issn.1673-8640.2024.06.013

摘要/Abstract

摘要：

目的探讨基于基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）的直接靶板微滴生长测定（DOT-MGA）法快速筛查碳青霉烯类耐药肺炎克雷伯菌（CRKP）的准确性和临床应用价值。方法收集2020—2021年连云港市第一人民医院肺炎克雷伯菌临床分离株251株，其中美罗培南敏感123株，CRKP 128株。采用DOT-MGA法对4和6 μL 2种取样量的所有菌株进行美罗培南耐药性检测。采用微量肉汤稀释法测定菌株的美罗培南最小抑菌浓度（MIC）。以微量肉汤稀释法耐药性分析结果为标准，评价DOT-MGA法耐药性检测结果的准确性。结果以微量肉汤稀释法为标准，孵育生长4 h后，DOT-MGA法2种取样量的敏感性和阴性预测值均为100.0%，其中4 μL取样量检测CRKP的特异性为92.5%，阳性预测值为92.8%；6 μL取样量检测CRKP的特异性为89.8%，阳性预测值为90.1%。123株美罗培南敏感株的美罗培南MIC₅₀为0.125 μg·mL^-1，MIC₉₀为0.25 μg·mL^-1；128株美罗培南耐药株的MIC₅₀和MIC₉₀均为≥128 μg·mL^-1。DOT-MGA法与微量肉汤稀释法的一致性分析结果显示，4 μL加样量Kappa值为0.88，6 μL加样量Kappa值为0.83。结论基于MALDI-TOF MS的DOT-MGA法筛查CRKP具有较高的准确性，且简便、快捷，可用于快速检测CRKP。

关键词: 碳青霉烯类耐药肺炎克雷伯菌, 基质辅助激光解吸电离飞行时间质谱, 直接靶板微滴生长测定法, 微量肉汤稀释法

Abstract:

Objective To investigate the accuracy and clinical application role of direct-on-target microdroplet growth assay（DOT-MGA） based on matrix-assisted laser desorption ionization time-of-flight mass spectrometry（MALDI-TOF MS） for rapid determination of carbapenem-resistant Klebsiella pneumoniae（CRKP）. Methods A total of 251 clinical Klebsiella pneumoniae isolates were collected from the First Hospital of Lianyungang from 2020 to 2021，which included 123 meropenem-sensitive isolates and 128 CRKP isolates. The carbapenem resistance of all the isolates was determined by DOT-MGA with 2 sample sizes of 4 and 6 μL，and the minimum inhibitory concentration（MIC） of meropenem of each isolate was determined by broth microdilution method. Using the results of broth microdilution method as reference，the accuracy of DOT-MGA drug resistance results was evaluated. Results With the broth microdilution method as reference，the sensitivity and negative predictive value of DOT-MGA were 100.0% after incubation and growth for 4 h，and the specificity and positive predictive value of 4 μL group were 92.5% and 92.8%，respectively. The specificity and positive predictive value of 6 μL group were 89.8% and 90.1%，respectively. The MIC₅₀ of 123 meropenem-sensitive isolates was 0.125 μg·mL^-1，and the MIC₉₀ was 0.25 μg·mL^-1. The MIC₅₀ of 128 meropenem-resistant isolates was ≥128 μg·mL^-1，and the MIC₉₀ was ≥128 μg·mL^-1 as well. The consistency analysis results of DOT-MGA and broth microdilution method showed that the Kappa value of 4 μL group was 0.88，and that of 6 μL group was 0.83. Conclusions DOT-MGA based on MALDI-TOF is a simple，rapid and accurate method for the determination of CRKP，which can be used in the rapid determination of CRKP in clinical practice.

Key words: Carbapenem-resistant Klebsiella pneumoniae, Matrix-assisted laser desorption ionization time-of-flight mass spectrometry, Direct-on-target microdroplet growth assay, Broth microdilution method

中图分类号:

R378.99

吴昕哲, 茆海丰, 杨晋, 左春磊, 金丹婷. MALDI-TOF MS直接靶板微滴生长测定法在CRKP快速检测中的应用[J]. 检验医学, 2024, 39(6): 587-591.

WU Xinzhe, MAO Haifeng, YANG Jin, ZUO Chunlei, JIN Danting. Clinical application of MALDI-TOF direct droplet culture method in rapid determination of CRKP[J]. Laboratory Medicine, 2024, 39(6): 587-591.

图/表 2

参考文献 14

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