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

Taylor A. Wahlig Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah;

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Ben J. Brintz Division of Epidemiology, Department of Internal Medicine, University of Utah School of Medicine, Salt Lake City, Utah;

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Melanie Prettyman Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah;

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Andrew S. Azman Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland;

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Daniel T. Leung Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah;
Division of Microbiology and Immunology, University of Utah School of Medicine, Salt Lake City, Utah

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ABSTRACT.

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.

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Author Notes

 Address correspondence to Daniel T. Leung, Division of Infectious Diseases University of Utah, 26 N Medical Dr., Wintrobe 513, Salt Lake City, UT 84132. E-mail: daniel.leung@utah.edu

Financial support: This research was supported by the National Institutes of Health under Ruth L. Kirschstein National Research Service Award 5T32HL105321 (to T.A.W.), by R01AI135115 (to D.T.L.), and by the University of Utah Study Design and Biostatistics Center, with funding in part from the National Center for Research Resources and the National Center for Advancing Translational Sciences, National Institute of Health through grant 8UL1TR000105 (to B.J.B.).

Authors’ addresses: Taylor A. Wahlig and Melanie Prettyman, Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, UT, E-mails: u6026936@utah.edu and melanie.prettyman@utah.edu. Ben J. Brintz, Division of Epidemiology, Department of Internal Medicine, University of Utah School of Medicine, Salt Lake City, UT, E-mail: ben.brintz@hsc.utah.edu. Andrew S. Azman, Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD, E-mail: azman@jhu.edu. Daniel T. Leung, Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, UT, and Division of Microbiology and Immunology, University of Utah Health, Salt Lake City, UT, E-mail: daniel.leung@utah.edu.

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