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评估实时直接分析质谱法在嗜肺军团菌的鉴定和血清分型中的应用。

Assessing direct analysis in real-time mass spectrometry for the identification and serotyping of Legionella pneumophila.

机构信息

Istituto Zooprofilattico Sperimentale delle Venezie, Laboratorio di Chimica Sperimentale, Vicenza, Italy.

Department of Food Safety, Istituto Zooprofilattico Sperimentale delle Venezie, Legnaro, Italy.

出版信息

J Appl Microbiol. 2022 Feb;132(2):1479-1488. doi: 10.1111/jam.15301. Epub 2021 Sep 28.

Abstract

AIMS

The efficacy of ambient mass spectrometry to identify and serotype Legionella pneumophila was assessed. To this aim, isolated waterborne colonies were submitted to a rapid extraction method and analysed by direct analysis in real-time mass spectrometry (DART-HRMS).

METHODS AND RESULTS

The DART-HRMS profiles, coupled with partial least squares discriminant analysis (PLS-DA), were first evaluated for their ability to differentiate Legionella spp. from other bacteria. The resultant classification model achieved an accuracy of 98.1% on validation. Capitalising on these encouraging results, DART-HRMS profiling was explored as an alternative approach for the identification of L. pneumophila sg. 1, L. pneumophila sg. 2-15 and L. non-pneumophila; therefore, a different PLS-DA classifier was built. When tested on a validation set, this second classifier reached an overall accuracy of 95.93%. It identified the harmful L. pneumophila sg. 1 with an impressive specificity (100%) and slightly lower sensitivity (91.7%), and similar performances were reached in the classification of L. pneumophila sg. 2-15 and L. non-pneumophila.

CONCLUSIONS

The results of this study show the DART-HMRS method has good accuracy, and it is an effective method for Legionella serogroup profiling.

SIGNIFICANCE AND IMPACT OF THE STUDY

These preliminary findings could open a new avenue for the rapid identification and quick epidemiologic tracing of L. pneumophila, with a consequent improvement to risk assessment.

摘要

目的

评估环境质谱法识别和血清分型嗜肺军团菌的效果。为此,将分离的水载菌落进行快速提取方法处理,并通过实时直接分析质谱法(DART-HRMS)进行分析。

方法和结果

首先,DART-HRMS 图谱结合偏最小二乘判别分析(PLS-DA),用于评估其区分嗜肺军团菌属与其他细菌的能力。所得分类模型在验证中达到了 98.1%的准确性。利用这些令人鼓舞的结果,探索了 DART-HRMS 图谱作为鉴定嗜肺军团菌 sg.1、嗜肺军团菌 sg.2-15 和非嗜肺军团菌的替代方法;因此,建立了不同的 PLS-DA 分类器。在验证集上进行测试时,该第二个分类器的总体准确性达到了 95.93%。它以 100%的出色特异性(特异性)和稍低的敏感性(91.7%)识别了有害的嗜肺军团菌 sg.1,并且在嗜肺军团菌 sg.2-15 和非嗜肺军团菌的分类中达到了类似的性能。

结论

本研究结果表明,DART-HMRS 方法具有良好的准确性,是一种有效的嗜肺军团菌血清分型方法。

研究的意义和影响

这些初步发现可能为快速鉴定和快速进行流行病学追踪嗜肺军团菌开辟了新途径,从而改善风险评估。

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