Mass Spectrometry–Based Escherichia coli H Antigen/Flagella Typing: Validation and Comparison with Traditional Serotyping
| Autor: | Keding Cheng, Aleisha Reimer, Bianli Xu, Larissa Domish, Gehua Wang, Angela Sloan, Jun Ma, J. David Knox, Yi-Min She, Huixia Chui, Drexler Hernandez, Stuart McCorrister |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Serotype Canada Clinical Biochemistry H antigen Biology medicine.disease_cause Sensitivity and Specificity 01 natural sciences Mass Spectrometry Microbiology 03 medical and health sciences Protein sequencing Escherichia coli medicine Typing Serotyping Whole genome sequencing Antiserum Antigens Bacterial 010401 analytical chemistry Biochemistry (medical) Repeatability Molecular biology 0104 chemical sciences 030104 developmental biology Flagella |
| Zdroj: | Clinical Chemistry. 62:839-847 |
| ISSN: | 1530-8561 0009-9147 |
| DOI: | 10.1373/clinchem.2015.244236 |
| Popis: | BACKGROUND Escherichia coli H antigen typing with antisera, a useful method for flagella clinical identification and classification, is a time-consuming process because of the need to induce flagella growth and the occurrence of undetermined strains. We developed an alternative rapid and analytically sensitive mass spectrometry (MS) method, termed MS-based H antigen typing (MS-H), and applied it at the protein sequence level for H antigen typing. We also performed a comparison with traditional serotyping on reference strains and clinical isolates. METHODS On the basis of international guidelines, the analytical selectivity and sensitivity, imprecision, correlation, repeatability, and reproducibility of the MS-H platform was evaluated using reference strains. Comparison of MS-H typing and serotyping was performed using 302 clinical isolates from 5 Canadian provinces, and discrepant results between the 2 platforms were resolved through whole genome sequencing. RESULTS Repeated tests on reference strain EDL933 demonstrated a lower limit of the measuring interval at the subsingle colony (16.97 μg or 1.465 × 107 cells) level and close correlation (r2 > 0.99) between cell culture biomass and sequence coverage. The CV was CONCLUSIONS MS-H typing of E. coli is useful for fast and accurate flagella typing and could be very useful during E. coli outbreaks. |
| Databáze: | OpenAIRE |
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