MALDI-TOF MS法测定拉脱维亚临床分离株中耐甲氧西林金黄色葡萄球菌和表皮葡萄球菌。

Pub Date : 2020-04-01 DOI:10.1016/j.clinms.2020.03.001
Laura Alksne, Svetlana Makarova, Jeļena Avsejenko, Alla Cibrovska, Julija Trofimova, Olga Valciņa
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引用次数: 6

摘要

快速鉴定耐甲氧西林葡萄球菌可确保适当的医疗护理。利用409株葡萄球菌,建立了可靠的MALDI-TOF方法进行菌种鉴定。以12株金黄色葡萄球菌为实验对象,比较了3种不同的制样方法和直接法测定耐甲氧西林m/z 2414±2指示峰的重复性。利用临床和医院环境分离的65株葡萄球菌(包括37株耐甲氧西林葡萄球菌)证实存在酚溶性调节素(PSM-mec)肽。409份样品的272株金黄色葡萄球菌均通过MALDI-TOF在种水平上正确鉴定。用三种方法制备的样品所得到的光谱在强度和某些峰的存在上存在差异。提取后的PSM-mec峰不可见。峰m/z 2414±2仅在61%的甲氧西林耐药菌株中检测到,未在甲氧西林敏感菌株中检测到。5株金黄色葡萄球菌在m/z 2414±2处的峰重现性为87%。无论甲氧西林敏感性如何,在65个样品中的49个样品以及所有显示PSM-mec峰的样品中均观察到delta-毒素。峰m/z 2414±2是携带mecA基因的耐甲氧西林菌株特有的,但峰m/z 2414±2的缺失并不排除对甲氧西林耐药的可能性。因此,在常规实验室工作中实施MALDI-TOF分析,特别是在临床样本中,在许多情况下可以提供关于耐甲氧西林菌株存在的快速警告。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Determination of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis by MALDI-TOF MS in clinical isolates from Latvia

Rapid identification of methicillin-resistant Staphylococcus could ensure appropriate medical care. A total of 409 Staphylococcus spp. strains were used to develop a reliable MALDI-TOF method for species identification. We tested twelve S. aureus strains to compare three different sample preparation methods and the reproducibility of the methicillin-resistant m/z 2414 ± 2 indicator peak with direct method in triplicate. A total of 65 Staphylococcus spp. strains (including 37 methicillin-resistant strains) from clinical and hospital environment isolates were used to confirm the presence of phenol-soluble modulin (PSM-mec) peptide. All 272 S. aureus strains from 409 samples were correctly identified at species level by MALDI-TOF. The samples prepared by three methods gave spectra with differences in the intensities and presence of certain peaks. The PSM-mec peak was not visible after the extraction method. The peak m/z 2414 ± 2 was only detected in 61% of the methicillin-resistant strains and in none of the methicillin-sensitive strains. The peak reproducibility for the five analyzed S. aureus strains showing the peak at m/z 2414 ± 2 was 87%. The delta-toxin was observed in 49 out of 65 samples regardless of methicillin susceptibility, as well as in all the samples exhibiting the PSM-mec peak. The peak m/z 2414 ± 2 is specific to methicillin-resistant strains carrying the mecA gene, but the absence of peak m/z 2414 ± 2 does not exclude the possibility of resistance to methicillin. Thus, implementing MALDI-TOF analysis in routine laboratory work, especially with clinical samples, would in many cases provide rapid warning about the presence of methicillin-resistant strains.

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