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一种基于基质辅助激光解吸/电离飞行时间质谱与流式细胞术相结合的快速鉴定病原体及药敏试验的新方法。

A New Method Aimed to Quickly Identify Pathogen and Drug Susceptibility Test Based on Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry Combined with Flow Cytometry.

作者信息

Gu Yu-Feng, Li Yu, Zhang Xiao-Lu, Yu Li-Mei, Huang Bao-Hua, Sun Cheng-Ming

机构信息

Department of Clinical Laboratory Center, Yantai Yuhuangding Hospital, Yantai, Shandong, China.

出版信息

Surg Infect (Larchmt). 2019 Apr;20(3):219-224. doi: 10.1089/sur.2018.145. Epub 2019 Jan 18.

DOI:10.1089/sur.2018.145
PMID:30657423
Abstract

AIM

To discuss the significance and applied value in the rapid identification and drug susceptibility test for blood stream infection (BSI) using matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) combined with flow cytometry (FCM).

METHODS

The bacteria were separated from the positive blood culture bottle using the separation gel-adsorption method system, and then applying MALDI-TOF MS combined with FCM to identify pathogen and drug susceptibility test quickly.

RESULTS

The efficiency of the separation gel-adsorption method for gram-negative bacterium, gram-positive bacteria, and fungi is 71%, 74%, and 88%, respectively. The results of identifying pathogens using MALDI-TOFMS are in agreement with results obtained using VITEK2 (bioMérieux, Marcy l'Etoile, France); both methods can identify 90% of bacteria to species. For fungi, MALDI-TOF MS can identify 75% fungi to species, which is superior to VITEK2, which identifies 60% fungi to species. The results of drug susceptibility test using FCM are almost identical to VITEK2; additionally, the addition of fluorescein diacetate can identify the heterogenic drug-resistant strains.

CONCLUSIONS

We can quickly identify pathogen and drug-susceptibility test based on MALDI-TOF MS combined with FCM, which is consistent with traditional methods and can shorten the report time from 36-72 hour to 3 hours. More importantly, these methods are of great significance and clinical importance for the rapid identification of BSI.

摘要

目的

探讨基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF MS)联合流式细胞术(FCM)在血流感染(BSI)快速鉴定及药敏试验中的意义和应用价值。

方法

采用分离胶吸附法系统从阳性血培养瓶中分离细菌,然后应用MALDI-TOF MS联合FCM快速鉴定病原体并进行药敏试验。

结果

分离胶吸附法对革兰阴性菌、革兰阳性菌和真菌的分离效率分别为71%、74%和88%。使用MALDI-TOF MS鉴定病原体的结果与使用VITEK2(法国生物梅里埃公司,马西伊图瓦勒)获得的结果一致;两种方法均可将90%的细菌鉴定到种。对于真菌,MALDI-TOF MS可将75%的真菌鉴定到种,优于VITEK2,后者能将60% 的真菌鉴定到种。使用FCM进行药敏试验的结果与VITEK2几乎相同;此外,添加二乙酸荧光素可鉴定异质性耐药菌株。

结论

基于MALDI-TOF MS联合FCM可快速鉴定病原体并进行药敏试验,与传统方法结果一致,可将报告时间从36 - 72小时缩短至3小时。更重要的是,这些方法对BSI的快速鉴定具有重要意义和临床价值。

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