Capsule Polysaccharide Quantification

The capsule polysaccharide of strains was extracted and quantified as previously described^{21 (link),50 (link)}. Briefly, 1 × 10⁹ colony forming units (CFU) bacteria were pelleted for capsule uronic acid extraction. Bacteria were mixed with 1% Zwittergent 3–14 detergent (Sangon Biotech, catalogue number A610552) in 100 mM citric acid (Sangon Biotech, catalogue number A610055). Then uronic acid was precipitated by ethanol (Sinopharm Chemical Reagent, catalogue number 10009218), incubated with 0.0125 M sodium tetraborate/sulfuric acid (tetraborate, Sigma-Aldrich, catalogue number 221732; sulfuric acid, Sinopharm Chemical Reagent, catalogue number 10021608) and 0.125% carbazole absolute ethanol (carbazole, Sangon Biotech, catalogue number A600269), finally measured at 530 nm and correlated to a standard curve of d-glucuronic acid.

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Hu F., Pan Y., Li H., Han R., Liu X., Ma R., Wu Y., Lun H., Qin X., Li J., Wang A., Zhou M., Liu B., Zhou Z, & He P. (2024). Carbapenem-resistant Klebsiella pneumoniae capsular types, antibiotic resistance and virulence factors in China: a longitudinal, multi-centre study. Nature Microbiology, 9(3), 814-829.

Publication 2024

citric acid tetraborate sulfuric acid

Corresponding Organization : National Institute for Parasitic Diseases

Other organizations : Huashan Hospital, Fudan University, Shanghai Jiao Tong University, Soochow University, Ruijin Hospital

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Variable analysis

independent variables

Bacterial strain (number of colony forming units)

dependent variables

Capsule uronic acid content

control variables

Zwittergent 3-14 detergent concentration (1%)
Citric acid concentration (100 mM)
Sodium tetraborate and sulfuric acid concentration (0.0125 M)
Carbazole concentration in absolute ethanol (0.125%)

controls

Positive control: D-glucuronic acid standard curve
Negative control: Not explicitly mentioned

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