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[使用直接基质辅助激光解吸电离飞行时间质谱法快速鉴定阳性血培养物]

[EXPRESS IDENTIFICATION OF POSITIVE BLOOD CULTURES USING DIRECT MALDI-TOF MASS SPECTROMETRY].

作者信息

Popov D A, Ovseenko S T, Vostrikova T Yu

出版信息

Anesteziol Reanimatol. 2015 Sep-Oct;60(5):71-5.

Abstract

OBJECTIVE

To evaluate the effectiveness of direct identification of pathogens of bacteremia by direct matrix assisted laser desorption ionization time-flight mass spectrometry (mALDI-TOF) compared to routine method.

MATERIAL AND METHODS

A prospective study included 211 positive blood cultures obtained from 116 patients (106 adults and 10 children, aged from 2 weeks to 77 years old in the ICU after open heart surgery. Incubation was carried out under aerobic vials with a sorbent for antibiotics Analyzer BacT/ALERT 3D 120 (bioMerieux, France) in parallel with the primary sieving blood cultures on solid nutrient media with subsequent identification of pure cultures using MALDI-TOF mass spectrometry analyzer Vitek MS, bioMerieux, France routine method), after appropriate sample preparation we carried out a direct (without screening) MALDI-TOF mass spectrometric study of monocomponental blood cultures (n = 201).

RESULTS

using a routine method in 211 positive blood cultures we identified 23 types of microorganisms (Staphylococcus (n = 87), Enterobacteria- ceae (n = 71), Enterococci (n = 20), non-fermentative Gram-negative bacteria (n = 18), others (n = 5). The average time of incubation of samples to obtain a signal of a blood culture growth was 16.2 ± 7.4 h (from 3.75 to 51 hours.) During the first 12 hours of incubation, growth was obtained in 32.4% of the samples, and on the first day in 92.2%. In the direct mass spectrometric analysis mnonocomponental blood cultures (n = 201) is well defined up to 153 species of the sample (76.1%), while the share of successful identification of Gram-negative bacteria was higher than that of Gram-positive (85.4 and 69, 1%, respectively p = 0.01). The high degree of consistency in the results of standard and direct method of identifying blood cultures using MALDI-TOF mass spectrometry (κ = 0.96, p < 0.001; the samples included in the calculation for which both option given result). Duration of the direct mass spectrometric analysis, including sample preparation, was no longer than 1 hour:

CONCLUSION

The method of direct MALDI-TOF mass spectrometry allows to significantly speed up the identification of blood cultures that may contribute as much as possible early appointment effective regimes of starting antibiotic therapy.

摘要

目的

评估与常规方法相比,直接基质辅助激光解吸电离飞行时间质谱(mALDI-TOF)直接鉴定菌血症病原体的有效性。

材料与方法

一项前瞻性研究纳入了从116例患者(106例成人和10例儿童,年龄从2周至77岁,均为心脏直视手术后入住重症监护病房)获取的211份阳性血培养物。在装有抗生素吸附剂的需氧瓶中,使用BacT/ALERT 3D 120分析仪(法国生物梅里埃公司)进行培养,并同时在固体营养培养基上对血培养物进行初步筛选,随后使用法国生物梅里埃公司的Vitek MS MALDI-TOF质谱分析仪对纯培养物进行鉴定(常规方法),在进行适当的样品制备后,我们对单组分血培养物(n = 201)进行了直接(无需筛选)MALDI-TOF质谱研究。

结果

使用常规方法在211份阳性血培养物中鉴定出23种微生物(葡萄球菌(n = 87)、肠杆菌科(n = 71)、肠球菌(n = 20)、非发酵革兰氏阴性菌(n = 18)、其他(n = 5))。获得血培养生长信号的样品平均孵育时间为16.2±7.4小时(3.75至51小时)。在孵育的前12小时内,32.4%的样品出现生长,第一天有92.2%出现生长。在直接质谱分析中(n = 201),单组分血培养物中多达153种样品(76.1%)得到明确鉴定,而革兰氏阴性菌的成功鉴定率高于革兰氏阳性菌(分别为85.4%和69.1%,p = 0.01)。使用MALDI-TOF质谱法对血培养物进行标准鉴定和直接鉴定的结果具有高度一致性(κ = 0.96,p < 0.001;计算中纳入的样品两种方法均给出结果)。包括样品制备在内的直接质谱分析持续时间不超过1小时。

结论

直接MALDI-TOF质谱法能够显著加快血培养物的鉴定速度,这可能有助于尽早确定有效的初始抗生素治疗方案。

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