Counting and Recognizing Single Bacterial Cells by a Lanthanide-Encoding Inductively Coupled Plasma Mass Spectrometric Approach
Counting and Recognizing Single Bacterial Cells by a Lanthanide-Encoding Inductively Coupled Plasma Mass Spectrometric Approach
复制标题
通过镧系元素编码电感耦合等离子体质谱方法计数和识别单个细菌细胞
DOI：
10.1021/acs.analchem.9b01130
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发表时间：
2019
影响因子：
7.4
通讯作者：
Wang Qiuquan
中科院分区：
文献类型：
--
作者：
Liang Yong;Liu Qian;Zhou Yang;Chen Shi;Yang Limin;Zhu Min;Wang Qiuquan
计数和识别单个细菌细胞对于细菌诱导疾病的诊断和细胞间变异性以及相关抗生素耐药机制的研究至关重要。为了更准确地分析单个细菌细胞，一直需要更高灵敏度和选择性的方法。我们首次报道了一种镧系元素编码的电感耦合等离子体（ICP）质谱法用于计数和识别单个细菌细胞。将非规范炔-d-丙氨酸（aDA）加入到大肠杆菌、金黄色葡萄球菌、单核细胞增生李斯特菌、志贺氏菌和副溶血弧菌等5种典型菌株中，aDA被代谢组装到肽聚糖层支持的细菌细胞壁中，然后用1，4，7，10-四氮杂环十二烷-1，4，7-三-乙酸-10-叠氮丙基乙基乙酰胺-铕络合物（azide-DOTA-Eu）对aDA进行后点击标记。这种Eu标记的细菌细胞可以被认为是Eu工程化颗粒，当通过单颗粒ICP质谱法（spICPMS）测定151/153 Eu时，相对于单个细菌细胞本身，其递送超过5个数量级的自信号放大结果。这种代谢组装的DA介导的Eu编码信号放大策略突破了spICPMS的检测限，并确保我们直接计数单个细菌细胞。所计数的菌株可通过其相应的镧系元素（Ln）编码的多克隆抗体（Ln = 139 La、141 Pr、142 Nd、152 Sm和160 Gd）同时识别，作为细菌的特异性鉴定（Ln-pAb-ID）。此外，所开发的方法被应用于显示遗传上相同的金黄色葡萄球菌在万古霉素和Ag纳米颗粒处理下的不同行为，表明这种镧系元素编码的spICPMS方法提供了一种新的方法来发现仍然模糊的细胞间变异性。
Counting and recognizing single bacterial cells are crucial to the diagnosis of bacterium-induced disease and study of cell-to-cell variability as well as the related antibiotic resistance mechanism. A higher sensitive and selective method has always been desired for a more accurate single bacterial cell analysis. We report a lanthanide-encoding inductively coupled plasma (ICP) mass spectrometric approach for counting and recognizing single bacterial cells for the first time. When noncanonicalalkyne-d-alanine (aDA) was added to five typical bacterial strains ofEscherichia coli,Staphylococcus aureus,Listeria monocytogenes,Shigella dysenteriae, andVibrio parahemolyticus,aDA was metabolically assembled into the peptidoglycan layer-supported bacterial cell wall followed by post-clickable europium-tagging with 1,4,7,10-tetraazacyclododecane-1,4,7-tris-acetic acid-10-azidopropyl ethylacetamide-europium complex (azide-DOTA-Eu). Such Eu-tagged bacterial cells can be deemed as Eu-engineered particles, delivering more than 5 orders of magnitude self-signal-amplification outcome relative to the single bacterial cells themselves when151/153Eu is determined by single particle ICP mass spectrometry (spICPMS). This metabolic assembly ofaDA mediated Eu-encoding signal amplification strategy breaks through the detection limit ofspICPMS and ensures that we directly count a single bacterial cell. The individual bacterial strains we counted can be simultaneously recognized through their corresponding lanthanide (Ln)-coded polyclonal antibody (Ln =139La,141Pr,142Nd,152Sm, and160Gd, respectively), serving as a specific bacterial identification (Ln-pAb-ID). Moreover, the developed approach was applied to show the different behavior between genetically identicalStaphylococcus aureusunder the treatments of vancomycin and Ag nanoparticles, demonstrating that such a lanthanide-encodingspICPMS approach provided a new way to discover still ambiguous cell-to-cell variability.