PubAg

Main content area

Counting and Recognizing Single Bacterial Cells by a Lanthanide-Encoding Inductively Coupled Plasma Mass Spectrometric Approach

Author:
Liang, Yong, Liu, Qian, Zhou, Yang, Chen, Shi, Yang, Limin, Zhu, Min, Wang, Qiuquan
Source:
Analytical chemistry 2019 v.91 no.13 pp. 8341-8349
ISSN:
1520-6882
Subject:
Escherichia coli, Listeria monocytogenes, Shigella dysenteriae, Staphylococcus aureus, Vibrio parahaemolyticus, antibiotic resistance, bacteria, cell walls, detection limit, mass spectrometry, nanosilver, peptidoglycans, polyclonal antibodies, vancomycin
Abstract:
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 noncanonical alkyne-d-alanine (aDA) was added to five typical bacterial strains of Escherichia coli, Staphylococcus aureus, Listeria monocytogenes, Shigella dysenteriae, and Vibrio 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 when ¹⁵¹/¹⁵³Eu is determined by single particle ICP mass spectrometry (spICPMS). This metabolic assembly of aDA mediated Eu-encoding signal amplification strategy breaks through the detection limit of spICPMS 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, and ¹⁶⁰Gd, respectively), serving as a specific bacterial identification (Ln-pAb-ID). Moreover, the developed approach was applied to show the different behavior between genetically identical Staphylococcus aureus under the treatments of vancomycin and Ag nanoparticles, demonstrating that such a lanthanide-encoding spICPMS approach provided a new way to discover still ambiguous cell-to-cell variability.
Agid:
6489172