Rapid identification of mycobacteria from positive MGIT broths of primary cultures by MALDI-TOF mass spectrometry.
Rapid identification of mycobacteria from positive MGIT broths of primary cultures by MALDI-TOF mass spectrometry.
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DOI：
10.1371/journal.pone.0192291
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Lee SS
中科院分区：
文献类型：
--
作者：
Huang TS;Lee CC;Tu HZ;Lee SS
快速鉴定分枝杆菌对于及时治疗和实施公共卫生措施非常重要。MGIT系统可确保快速检测分枝杆菌，但由于在固体培养基上进一步传代培养，鉴定通常会延迟数天至数周。基质辅助激光解吸飞行时间质谱（MALDI-TOF MS）被证明可以有效地鉴定从固体或液体培养基中传代培养的分枝杆菌分离株。忽略传代培养步骤，直接从无菌和非无菌临床标本的MGIT肉汤中进行鉴定的报告有限，并且之前并不令人满意。我们的鉴定方法大大缩短了从检测到鉴定分枝杆菌的延迟。我们评估了Vitek MS IVD 3.0版用于直接鉴定原代MGIT培养物中NTM和结核分枝杆菌的性能，并评估了两种样品制备方法。使用MALDI-TOF VITEK MS（IVD v.3.0）直接鉴定阳性MGIT肉汤中的NTM，生成的可接受结果率较高，达到96.4%（80/83），包括0.1% SDS洗涤步骤的样品制备液的可接受结果率高达100%（83/83）。以免疫层析（伊卡）试验为金标准，VITEK MS鉴定MGIT管中结核分枝杆菌的灵敏度为58/72（80.6%）。在48例中观察到特征性的菌落聚集，羊毛样外观，所有58例（100%）均使用MALDI-TOF正确鉴定为结核分枝杆菌。24支MGIT管中结核分枝杆菌复合菌检出率较低（10/24，41.6%），为多菌性。我们的方法显著降低了试剂成本和周转时间。基于一个简化的协议，我们表明，MALDI-TOF MS可用于快速鉴定NTM直接从原代MGIT培养物在常规的临床实验室工作流程。然而，由于结核分枝杆菌的识别率较低，我们建议最初进行伊卡检测以筛查结核分枝杆菌复合群。使用MALDI-TOF在存在多微生物培养物的情况下检测结核病。
Rapid identification of mycobacteria is important for timely treatment and the implementation of public health measures. The MGIT system ensures rapid detection of mycobacteria, but identification is usually delayed by days to weeks due to further subculture on solid medium. Matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-TOF MS) was demonstrated to effectively identify mycobacteria isolates subcultured from solid or liquid media. Reports of identification directly from MGIT broths of both sterile and non-sterile clinical specimens, omitting the subculture step, were limited and not satisfactory before. Our identification method dramatically shortened delay from detection to identification of mycobacteria. We assessed the performance of the Vitek MS IVD version 3.0 for direct identification of NTM and M.tuberculosis from primary MGIT cultures, and assessed two sample preparation methods. Direct identification of NTM from positive MGIT broths, using MALDI-TOF VITEK MS with IVD v.3.0, generated high rates of acceptable results reaching 96.4% (80/83), and up to 100% (83/83) for sample preparations including a 0.1% SDS washing step. The sensitivity of VITEK MS to identify M.tuberculosis from MGIT tubes was 58/72 (80.6%), when using immunochromatography (ICA) test as gold standard. A characteristic colony clumping, wool-like appearance was observed in 48, and all 58 (100%) were correctly identified as M.tuberculosis using MALDI-TOF. The detection rate of M.tuberculosis complex was low (10/24, 41.6%) in the 24 MGIT tubes that was polymicrobial. Our method significantly reduced both the reagent cost and turnaround time. Based on a simplified protocol, we showed that MALDI-TOF MS can be used for rapid identification of NTM directly from primary MGIT cultures within the routine clinical laboratory workflow. However, we recommend an initial ICA test to screen for M.tuberculosis complex, due to a low identification rate of M. tuberculosis in the presence of polymicrobial cultures using MALDI-TOF.
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影响因子：
9.4
作者：
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影响因子：
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DOI：
10.1016/j.diagmicrobio.2010.12.011
发表时间：
2011-05-01
影响因子：
2.9
作者：
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通讯作者：
Soler, Charles