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Potential of MALDI-TOF MS as an alternative approach for capsular typing Streptococcus pneumoniae isolates

机译:MALDI-TOF MS作为荚膜分型肺炎链球菌分离株的替代方法的潜力

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摘要

Streptococcus pneumoniae can be classified in more than 90 capsular types, as traditionally determined by serological methods and more recently by PCR-based techniques. Such methods, however, can be expensive, laborious or unable to accurately discriminate among certain serotypes. Therefore, determination of capsular types, although extremely important for epidemiological purposes and for estimating the impact of pneumococcal conjugate vaccines, is mainly restricted to research laboratories, being rarely performed in the clinical setting. In the present study, MALDI-TOF MS was evaluated as an alternative tool to characterize 416 pneumococcal isolates belonging to serotypes 6A, 6B, 6C, 9N, 9V or 14. For MALDI-TOF MS analysis, each isolate was submitted to an extraction protocol using formic acid and acetonitrile. Measurements were performed with a Bruker Microflex LT mass spectrometer using default parameters and generating spectra in the range of 2,000–20,000 m/z. Spectra were analyzed with the BioNumerics software v7.6. Isolates were mainly distributed according to the capsular type in a Neighbor Joining tree and serotypes investigated were successfully discriminated by the presence/absence of 14 selected biomarkers. The results suggest that MALDI-TOF MS is a promising alternative for typing pneumococcal strains, highlighting its usefulness for rapid and cost-effective routine application in clinical laboratories.
机译:肺炎链球菌可以分为90种以上的荚膜类型,传统上是通过血清学方法确定的,最近是通过基于PCR的技术确定的。然而,这种方法可能是昂贵的,费力的或无法准确地区分某些血清型。因此,尽管荚膜类型的确定对于流行病学目的和评估肺炎球菌结合疫苗的影响极为重要,但主要限于研究实验室,在临床环境中很少进行。在本研究中,评估了MALDI-TOF MS作为表征416种血清型6A,6B,6C,9N,9V或14的肺炎球菌分离株的替代工具。对于MALDI-TOF MS分析,每种分离株均需进行提取操作用甲酸和乙腈。使用默认参数在Bruker Microflex LT质谱仪上进行测量,并生成2,000–20,000 m / z范围内的光谱。用BioNumerics软件v7.6分析光谱。分离物主要根据荚膜类型分布在邻居连接树中,并且通过14种选择的生物标志物的存在/不存在成功地区分了所研究的血清型。结果表明,MALDI-TOF MS是用于分型肺炎球菌菌株的一种有前途的替代方法,突显了其在临床实验室中快速且经济高效的常规应用的有用性。

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