Suwantarat Nuntra, Weik Chelsea, Romagnoli Mark, Ellis Brandon C, Kwiatkowski Nicole, Carroll Karen C
From the Division of Medical Microbiology,
Microbiology Laboratory, Johns Hopkins Hospital, Baltimore, MD.
Am J Clin Pathol. 2016 Jan;145(1):22-8. doi: 10.1093/ajcp/aqv006.
Corynebacterium species and gram-positive coryneform-like bacteria (coryneforms) are increasingly reported causes of opportunistic infections in immunocompromised patients. Biochemical identification methods for these bacteria are often inaccurate. We evaluated matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) for routine Corynebacterium and coryneform identification.
A total of 286 Corynebacterium species and coryneforms recovered from patients were identified by MALDI-TOF MS analysis using the Bruker Microflex instrument, Biotyper software version 3.0, and database version 3.1.66 (Bruker Daltonics, Billerica, MA) following formic acid-based, direct on-plate extraction. The spectral cutoff scores used for interpretation were 2.0 or more for species-level identification and 1.7 or more for genus level. Scores lower than 1.7 were considered as no reliable identification. The results were compared with API Coryne (bioMérieux, Durham, NC) and sequencing of 16S ribosomal RNA genes.
Of the 231 Corynebacterium (19 species), 99.6% were correctly identified to the genus level and 88.7% to the species level. Of the 55 coryneforms (14 genera), 90.9% were correctly identified to the genus level and 67.3% to the species level. API Coryne was able to identify 89.2% of Corynebacterium species (species level) and 63.6% of coryneforms (genus level).
Rapid on-plate testing yielded identification of more Corynebacterium species and related bacteria than biochemical methods.
棒状杆菌属细菌和革兰氏阳性棒状杆菌样细菌(棒状杆菌)越来越多地被报道为免疫功能低下患者机会性感染的病因。这些细菌的生化鉴定方法往往不准确。我们评估了基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)用于常规棒状杆菌和棒状杆菌样细菌鉴定的效果。
使用布鲁克Microflex仪器、Biotyper软件3.0版本和数据库3.1.66版本(布鲁克道尔顿公司,马萨诸塞州比勒里卡),通过基于甲酸的直接平板提取法,对从患者身上分离出的286株棒状杆菌属细菌和棒状杆菌样细菌进行MALDI-TOF MS分析鉴定。用于结果判读的光谱截断分数为:种水平鉴定为2.0及以上,属水平鉴定为1.7及以上。低于1.7的分数被认为鉴定不可靠。将结果与API Coryne(bioMérieux公司,北卡罗来纳州达勒姆)以及16S核糖体RNA基因测序结果进行比较。
在231株棒状杆菌(19个种)中,99.6%在属水平被正确鉴定,88.7%在种水平被正确鉴定。在55株棒状杆菌样细菌(14个属)中,90.9%在属水平被正确鉴定,67.3%在种水平被正确鉴定。API Coryne能够鉴定89.2%的棒状杆菌属细菌(种水平)和63.6%的棒状杆菌样细菌(属水平)。
与生化方法相比,快速平板检测能够鉴定出更多的棒状杆菌属细菌及相关细菌。