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使用FastPrep-2珠磨系统和基质辅助激光解吸电离飞行时间质谱技术从液体培养基(MGIT)中直接鉴定分枝杆菌菌种:与固体培养基和基于聚合酶链反应的方法的比较

Direct identification of Mycobacterium species from liquid medium (MGIT) using FastPrep-2 bead beating system and MALDI-TOF mass spectrometry technology: a comparison with solid media and PCR-based method.

作者信息

Bazzi Ali M, Sunki Abdulaziz A, Hamdi Mohab J, Khaldi Abdullah O, Al-Tawfiq Jaffar A

机构信息

Microbiology Laboratory, Johns Hopkins Aramco Healthcare, Dhahran, Saudi Arabia.

Infectious Disease Unit, Specialty Internal Medicine, Johns Hopkins Aramco Healthcare, Dhahran, Saudi Arabia.

出版信息

Pract Lab Med. 2025 May 30;46:e00479. doi: 10.1016/j.plabm.2025.e00479. eCollection 2025 Sep.

Abstract

BACKGROUND

Mycobacterial infections present significant global health challenges. Traditional diagnostic methods are inadequate for the identification of Mycobacterium species, highlighting the need for more efficient techniques. Matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) technology offers potential advantages in rapid and accurate pathogen identification.

OBJECTIVE

This study evaluates the accuracy and precision of the MALDI-TOF using the FastPrep-2 bead beating system and VITEK MS version 3.2 for identifying Mycobacterium species directly from Mycobacteria Growth Indicator Tube (MGIT), liquid medium, compared to MALDI-TOF (VITEK MS) based on the traditional solid medium (Lowenstein-Jensen). We also compared the result of the MALDI-TOF from MGIT to results by PCR-based method.

METHODS

The study included 16 (MTB) and 37 (NTM). Isolates were grown in LJ solid medium and MGIT liquid medium, and lysed using the FastPrep-2 bead beating system. Identification was performed using VITEK MS version 3.2 from liquid.

RESULTS

NTM comprised 70 % (37/53) of the total isolates evaluated. Overall, the method achieved 88.6 % accuracy for identifying species from liquid medium, compared to 96.2 % from solid medium. The agreement between both methods was moderate (Kappa = 0.470, p < 0.001). MTB isolates were identified with 100 % accuracy, and the approach demonstrated excellent reproducibility with 100 % intra-assay and inter-day consistency.

CONCLUSION

Using VITEK MS version 3.2 to directly identify MTB and NTM from MGIT liquid medium provides a rapid, cost-effective, and reliable method for identifying Mycobacterium species. Further optimization and database expansion are recommended to enhance accuracy for rare and less common mycobacterial species.

摘要

背景

分枝杆菌感染给全球健康带来了重大挑战。传统诊断方法在分枝杆菌菌种鉴定方面存在不足,这凸显了对更高效技术的需求。基质辅助激光解吸电离飞行时间(MALDI-TOF)质谱(MS)技术在快速准确鉴定病原体方面具有潜在优势。

目的

本研究评估了使用FastPrep-2珠磨系统和VITEK MS 3.2版本的MALDI-TOF直接从分枝杆菌生长指示管(MGIT)液体培养基中鉴定分枝杆菌菌种的准确性和精密度,并与基于传统固体培养基(洛温斯坦-詹森培养基)的MALDI-TOF(VITEK MS)进行比较。我们还将MGIT中MALDI-TOF的结果与基于PCR方法的结果进行了比较。

方法

该研究纳入了16株结核分枝杆菌(MTB)和37株非结核分枝杆菌(NTM)。分离株在LJ固体培养基和MGIT液体培养基中培养,并使用FastPrep-2珠磨系统进行裂解。使用VITEK MS 3.2版本从液体中进行鉴定。

结果

NTM占评估的总分离株的70%(37/53)。总体而言,该方法从液体培养基中鉴定菌种的准确率为88.6%,而从固体培养基中鉴定的准确率为96.2%。两种方法之间的一致性为中等(Kappa = 0.470,p < 0.001)。MTB分离株的鉴定准确率为100%,该方法在实验内和日间一致性均为100%,显示出极好的可重复性。

结论

使用VITEK MS 3.2版本直接从MGIT液体培养基中鉴定MTB和NTM为分枝杆菌菌种鉴定提供了一种快速、经济高效且可靠的方法。建议进一步优化和扩展数据库,以提高对罕见和不太常见分枝杆菌菌种的鉴定准确性。

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