Capillary electrophoresis as a method to determine underivatized urinary lipoarabinomannans, a biomarker of active tuberculosis caused by Mycobacterium tuberculosis

Tuberculosis is a devastating contagious disease caused by Mycobacterium tuberculosis. This is the first report describing the development of novel capillary electrophoresis methods to detect lipoarabinomannans shed into the blood circulation by replicating bacteria. The novelty of the methods is th...

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Bibliographic Details
Published in:Journal of separation science 2016-07, Vol.39 (14), p.2853-2861
Main Authors: Sirén, Heli, Savolainen, Laura E., Tuuminen, Tamara
Format: Article
Language:English
Subjects:
Absorbance
Bacteria
Biomarkers
Biomarkers - urine
Capillary electrophoresis
Dynamic range
Electrolytes - chemistry
Electrophoresis
Electrophoresis, Capillary
Humans
Ionization
Lipoarabinomannan
Lipopolysaccharides - urine
Mycobacterium tuberculosis
Mycobacterium tuberculosis - isolation & purification
Oligosaccharides
Polarity
Sheds
Tuberculosis
Tuberculosis - urine
Urine
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Summary:Tuberculosis is a devastating contagious disease caused by Mycobacterium tuberculosis. This is the first report describing the development of novel capillary electrophoresis methods to detect lipoarabinomannans shed into the blood circulation by replicating bacteria. The novelty of the methods is the detection without derivatization. The lipoarabinomannan is detected owing to the ionization of the diverse functional groups of the structure, such as the multibranched mannan domain or the phosphatidyl group. Four alkaline solutions were used; normal polarity in three of them and reversed polarity in one. Urinary lipoarabinomannans by saccharide domains were identified with direct absorbance detection. The accuracy and the analytical sensitivity were then validated with cello‐, manno‐ and xylooligosaccharides. Lipoarabinomannan detection was feasible within 20 min (RSD 2.1%). This method worked at the dynamic range of 0.1–10 μg/mL. With reversed polarity, indirect absorbance detection, and pH 9.0 electrolyte were used, the analytes migrated already within 5 min (RSD 0.01%). Inorganic nonabsorbing ions were used for this method optimization. This improvement resulted in the detection limit of 1 pg/mL in water and in the linear dynamic range of 1 pg/mL to 10 ng/mL. In conclusion, the described method has great potential as a point‐of‐care assay for clinical use.
ISSN:1615-9306
1615-9314
DOI:10.1002/jssc.201600166