| 摘要: |
| ［摘要］ 目的 建立基于核酸质谱技术快速鉴定肺炎克雷伯菌碳青霉烯酶基因型的方法。方法 收集2021年1月至2023年9月徐州医科大学附属医院分离的56株非重复耐碳青霉烯类肺炎克雷伯菌（CRKP），通过基质辅助激光解吸电离飞行时间质谱（MALDI-TOF MS）技术和VITEK-2 Compact完成菌株鉴定及药敏分析。设计特异性单碱基延伸引物，结合多重聚合酶链反应（PCR）扩增与核酸质谱技术，在同一反应体系中检测碳青霉烯酶基因型特征峰，分析核酸质谱结果与PCR结果的一致性。结果 56株CRKP中KPC、NDM、OXA-232及KPC+NDM联产型菌株各14株。各亚型特征峰无交叉干扰；核酸质谱对KPC、OXA-232及KPC+NPM联产耐药菌株检测结果与PCR结果的一致率为100.00%，仅误将1株NDM菌株划分为NDM+OXA联产型菌株，总体检测一致率为98.21%（55/56）。药敏试验结果显示，KPC及NDM型菌株对亚胺培南耐药率为100.00%，而OXA-232菌株对亚胺培南耐药率为0。结论 核酸质谱技术可高效、精准鉴定CRKP碳青霉烯酶基因型，与PCR结果高度一致，为临床快速分型及耐药监测提供了可靠工具。 |
| 关键词: 肺炎克雷伯菌 碳青霉烯酶 基质辅助激光解吸电离飞行时间质谱 核酸质谱技术 |
| DOI：10.3969/j.issn.1674-3806.2025.07.03 |
| 分类号:R 378 |
| 基金项目:江苏省卫生健康委科研项目（编号：Z2021009，Ym2023110）；徐州市科技项目（编号：KC23269） |
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| A study on application value of mass spectrometry nucleic acid detection technology in rapid identification of carbapenemase genotypes in Klebsiella pneumoniae |
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YE Lin1, LIANG Xinyi1, DENG Xinyi1, ZHAO Shulong2, SONG Shuang2, SUN Jingfang2, KANG Haiquan2
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1.School of Medical Technology, Xuzhou Medical University, Xuzhou 221004, China; 2.Department of Laboratory Medicine, the Affiliated Hospital of Xuzhou Medical University, Xuzhou 221002, China
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| Abstract: |
| ［Abstract］ Objective To establish a method for rapid identification of carbapenemase genotypes in Klebsiella pneumoniae on the basis of mass spectrometry nucleic acid detection technology. Methods A total of 56 non-redundant strains of carbapenem-resistant Klebsiella pneumoniae(CRKP) isolated in the Affiliated Hospital of Xuzhou Medical University from January 2021 to September 2023 were collected. Bacterial identification and antimicrobial susceptibility analysis were performed using matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry(MALDI-TOF MS) and VITEK-2 Compact system. Specific single-base extension primers were designed, and a multiplex polymerase chain reaction(PCR) combined with mass spectrometry nucleic acid detection technology was employed to detect the characteristic peaks of carbapenemase genotypes in a single reaction system. The consistency of the results of identifying the carbapenemase genotypes in CRKP by using mass spectrometry nucleic acid detection technology and using PCR was analyzed. Results Among the 56 strains of CRKP, there were 14 strains with KPC, 14 strains with NDM, 14 strains with OXA-232, and 14 strains with co-production of KPC+NDM. There was no cross-interference among the characteristic peaks of each subtype. The consistency between the detection results using mass spectrometry nucleic acid detection technology and using PCR for KPC-, OXA-232- and (KPC+NDM)- resistant strains was 100.00%. Only 1 strain with NDM was mistakenly classified as NDM+OXA co-production strain, and the overall detection consistency rate was 98.21%(55/56). The results of the drug sensitivity test showed that the resistance rate of the strains with KPC or the strains with NDM to imipenem was 100.00%, while the resistance rate of the strains with OXA-232 to imipenem was 0. Conclusion The mass spectrometry nucleic acid detection technology can efficiently and accurately identify the carbapenemase genotypes in CRKP, which is highly consistent with the results of PCR. This mass spectrometry nucleic acid detection technology provides a reliable tool for rapid clinical typing and drug resistance monitoring. |
| Key words: Klebsiella pneumoniae Carbapenemase Matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry(MALDI-TOF MS) Mass spectrometry nucleic acid detection technology |
