A Novel Luminescence-Based Serum Bactericidal Assay for Vibrio cholerae Reduces Assay Variation, Is Time- and Cost-Effective, and Directly Measures Continuous Titer Values.

Autor: Wahlig TA; 1Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah., Brintz BJ; 2Division of Epidemiology, Department of Internal Medicine, University of Utah School of Medicine, Salt Lake City, Utah., Prettyman M; 1Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah., Azman AS; 3Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland., Leung DT; 1Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah.; 4Division of Microbiology and Immunology, University of Utah School of Medicine, Salt Lake City, Utah.
Jazyk: angličtina
Zdroj: The American journal of tropical medicine and hygiene [Am J Trop Med Hyg] 2021 Jul 08; Vol. 105 (3), pp. 622-626. Date of Electronic Publication: 2021 Jul 08.
DOI: 10.4269/ajtmh.21-0136
Abstrakt: Cholera remains a significant public health burden worldwide, and better methods for monitoring cholera incidence would enhance the effectiveness of public health interventions. The serum bactericidal assay (SBA) has been used extensively for Vibrio cholerae vaccine assessments and serosurveillance. Current SBA approaches for V. cholerae rely on colony enumeration or optical density (OD600nm) readings to measure viable bacteria following complement-mediated lysis. These methods provide titer values that are constrained to discrete dilution values and rely on bacterial outgrowth, which is time consuming and prone to variation. Detection of bacterial proteins following complement-mediated lysis presents a faster and potentially less variable alternative approach independent of bacterial outgrowth. Here, we present an SBA that measures luciferase luminescence driven by lysis-released adenylate kinase. This approach is faster and less variable than growth-dependent SBAs and directly measures continuous titer values. This novel SBA method can potentially be applied to other bacteria of interest.
Databáze: MEDLINE
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