- ABSTRACT.
- INTRODUCTION
- MATERIALS AND METHODS
- RESULTS
- Impact of incubation time on titer in OD600nm SBA.
- The luminescence-based SBA has lower inter- and intra-assay variability and directly measures continuous titer values.
- The luminescence SBA can be used for field-collected and pre- and postvaccine sera and is complement specific.
- The luminescence SBA is more time- and cost-effective.
- DISCUSSION
- ACKNOWLEDGMENTS
- References
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Boyd MA, Tennant SM, Saague VA, Simon R, Muhsen K, Ramachandran G, Cross AS, Galen JE, Pasetti MF, Levine MM, 2014. Serum bactericidal assays to evaluate typhoidal and nontyphoidal Salmonella vaccines. Clin Vaccine Immunol 21: 712–721.
-
2.
Corbeil LB, Wunderlich AC, Ito JI, McCutchan AJ, 1978. Plaque assay for measuring serum bactericidal activity against gonococci. J Clin Microbiol 8: 618–620.
-
3.
Martin D, McCallum L, Glennie A, Ruijne N, Blatchford P, O’Hallahan J, Oster P, 2005. Validation of the serum bactericidal assay for measurement of functional antibodies against group B meningococci associated with vaccine trials. Vaccine 23: 2218–2221.
-
4.
Muschel LH, Treffers HP, 1956. Quantitative studies on the bactericidal actions of serum and complement: I. a rapid photometric growth assay for bactericidal activity. J Immunol 76: 1–10.
-
5.
Townsend K, Ladhani SN, Findlow H, Borrow R, 2014. Evaluation and validation of a serum bactericidal antibody assay for Haemophilus influenzae type b and the threshold of protection. Vaccine 32: 5650–5656.
-
6.
Attridge SR, Qadri F, Albert MJ, Manning PA, 2000. Susceptibility of Vibrio cholerae O139 to antibody-dependent, complement-mediated bacteriolysis. Clin Diagn Lab Immunol 7: 444–450.
-
7.
Jang MS, Sahastrabuddhe S, Yun C, Han SH, Yang JS, 2016. Serum bactericidal assay for the evaluation of typhoid vaccine using a semi-automated colony-counting method. Microb Pathog 97: 19–26.
-
8.
Liu X, Wang S, Sendi L, Caulfield MJ, 2004. High-throughput imaging of bacterial colonies grown on filter plates with application to serum bactericidal assays. J Immunol Methods 292: 187–193.
-
9.
Nahm MH, Yu J, Weerts HP, Wenzel H, Tamilselvi SC, Chandrasekaran L, Pasetti MF, Mani S, Kaminski RW, 2018. Development, Interlaboratory evaluations, and application of a simple, high-throughput Shigella serum bactericidal assay. mSphere 3: 1–14.
-
10.
Weerts HP, Yu J, Kaminski RW, Nahm MHA, 2019. A high-throughput Shigella-specific bactericidal assay. J Vis Exp 144: 1–9.
-
11.
Yang JS, Kim HJ, Yun C, Kang S, Im J, Kim H, Han SH, 2007. A semi-automated vibriocidal assay for improved measurement of cholera vaccine-induced immune responses. J Microbiol Methods 71: 141–146.
-
12.
Ayalew S, Confer AW, Shrestha B, Payton ME, 2012. A rapid microtiter plate serum bactericidal assay method for determining serum complement-mediated killing of Mannheimia haemolytica. J Microbiol Methods 89: 99–101.
-
13.
Sharma SK, Fatma T, Thukral SS, 1999. A simple and rapid serum bactericidal assay and its evaluation in clinical isolates of Klebsiella pneumoniae. J Microbiol Methods 39: 45–48.
-
14.
Rouphael NG, Satola S, Farley MM, Rudolph K, Schmidt DS, Gomez-de-leo P, Robbins JB, Schneerson R, Carlone GM, Romero-Steiner S, 2011 . Evaluation of serum bactericidal antibody assays for Haemophilus influenzae serotype a. Clin Vaccine Immunol 18: 243–247.
-
15.
Mountzouros KT, Howell AP, 2000. Detection of complement-mediated antibody-dependent bactericidal activity in a fluorescence-based serum bactericidal assay for group B Neisseria meningitidis. J Clin Microbiol 38: 2878–2884.
-
16.
Romero-Steiner S, Spear W, Brown N, Holder P, Hennessy T, de Leon PG, Carlone GM, 2004. Measurement of serum bactericidal activity specific for Haemophilus influenzae type b by using a chromogenic and fluorescent metabolic indicator. Clin Diagn Lab Immunol 11: 89–93.
-
17.
Rodriguez T, Lastre M, Cedre B, del Campo J, Bracho G, Zayas C, Taboada C, Diaz M, Sierra G, Perez O, 2002. Standardization of Neisseria meningitidis serogroup B colorimetric serum bactericidal assay. Clin Diagn Lab Immunol 9: 109–114.
-
18.
Necchi F, Saul A, Rondini S, 2017. Development of a high-throughput method to evaluate serum bactericidal activity using bacterial ATP measurement as survival readout. PLoS One 12: e0172163.
-
19.
Necchi F, Saul A, Rondini S, 2018. Setup of luminescence-based serum bactericidal assay against Salmonella Paratyphi A. J Immunol Methods 461: 117–121.
-
20.
Rossi O, Molesti E, Saul A, Giannelli C, Micoli F, Necchi F, 2020. Intra-laboratory evaluation of luminescence based high-throughput serum bactericidal assay (L-SBA) to determine bactericidal activity of human sera against Shigella .High Throughput 9: 1–12.
-
21.
Clow F, O’Hanlon CJ, Christodoulides M, Radcliff FJ, 2019. Feasibility of using a luminescence-based method to determine serum bactericidal activity against Neisseria gonorrhoeae. Vaccines (Basel) 7: 191.
-
22.
Jacobs AC, Didone L, Jobson J, Sofia MK, Krysan D, Dunman M, 2013. Adenylate kinase release as a high-throughput-screening-compatible reporter of bacterial lysis for identification of antibacterial agents. Antimicrob Agents Chemother 57: 26–36.
-
23.
Dzeja P, Terzic A, 2009. Adenylate kinase and AMP signaling networks: metabolic monitoring, signal communication and body energy sensing. Int J Mol Sci 10: 1729–1772.
-
24.
Ali M, Nelson AR, Lopez AL, Sack DA, 2015. Updated global burden of cholera in endemic countries. PLoS Negl Trop Dis 9: e0003832.
-
25.
Attridge SR, Johansson C, Trach DD, Qadri F, 2002. Sensitive microplate assay for detection of bactericidal antibodies to Vibrio cholerae O139. Clin Diagn Lab Immunol 9: 383–387.
-
26.
Azman AS et al., 2019. Estimating cholera incidence with cross-sectional serology. Sci Transl Med 11: 1–11.
-
27.
Richie E et al., 2000. Efficacy trial of single-dose live oral cholera vaccine CVD 103-HgR in North Jakarta, Indonesia, a cholera-endemic area. Vaccine 18: 2399–2410.
-
28.
Tacket CO et al., 1999. Randomized, double-blind, placebo-controlled, multicentered trial of the efficacy of a single dose of live oral cholera vaccine CVD 103-HgR in preventing cholera following challenge with Vibrio cholerae O1 El Tor Inaba three months after vaccination. Infect Immun 67: 6341–6345.
-
29.
Boutonnier A, Dassy B, Dumenil R, Guenole A, Ratsitorahina M, Migliani R, Fournier J, 2003. A simple and convenient microtiter plate assay for the detection of bactericidal antibodies to Vibrio cholerae O1 and Vibrio cholerae O139. J Microbiol Methods 55: 745–753.
-
30.
Kauffman RC et al., 2016. Single-cell analysis of the plasmablast response to Vibrio cholerae demonstrates expansion of cross-reactive memory B cells. mBio 7: 1–11.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1147 | 555 | 37 |
| Full Text Views | 207 | 12 | 0 |
| PDF Downloads | 159 | 14 | 0 |
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
Taylor A. Wahlig
Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah;
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Ben J. Brintz
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
Melanie Prettyman
Division of Infectious Diseases, University of Utah School of Medicine, Salt Lake City, Utah;
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Andrew S. Azman
Andrew S. Azman
Department of Epidemiology, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland;
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Daniel T. Leung
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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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.
- Supplemental Materials (PDF 849 KB)
Author Notes
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.
- ABSTRACT.
- INTRODUCTION
- MATERIALS AND METHODS
- RESULTS
- Impact of incubation time on titer in OD600nm SBA.
- The luminescence-based SBA has lower inter- and intra-assay variability and directly measures continuous titer values.
- The luminescence SBA can be used for field-collected and pre- and postvaccine sera and is complement specific.
- The luminescence SBA is more time- and cost-effective.
- DISCUSSION
- ACKNOWLEDGMENTS
- References
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
Boyd MA, Tennant SM, Saague VA, Simon R, Muhsen K, Ramachandran G, Cross AS, Galen JE, Pasetti MF, Levine MM, 2014. Serum bactericidal assays to evaluate typhoidal and nontyphoidal Salmonella vaccines. Clin Vaccine Immunol 21: 712–721.
-
2.
Corbeil LB, Wunderlich AC, Ito JI, McCutchan AJ, 1978. Plaque assay for measuring serum bactericidal activity against gonococci. J Clin Microbiol 8: 618–620.
-
3.
Martin D, McCallum L, Glennie A, Ruijne N, Blatchford P, O’Hallahan J, Oster P, 2005. Validation of the serum bactericidal assay for measurement of functional antibodies against group B meningococci associated with vaccine trials. Vaccine 23: 2218–2221.
-
4.
Muschel LH, Treffers HP, 1956. Quantitative studies on the bactericidal actions of serum and complement: I. a rapid photometric growth assay for bactericidal activity. J Immunol 76: 1–10.
-
5.
Townsend K, Ladhani SN, Findlow H, Borrow R, 2014. Evaluation and validation of a serum bactericidal antibody assay for Haemophilus influenzae type b and the threshold of protection. Vaccine 32: 5650–5656.
-
6.
Attridge SR, Qadri F, Albert MJ, Manning PA, 2000. Susceptibility of Vibrio cholerae O139 to antibody-dependent, complement-mediated bacteriolysis. Clin Diagn Lab Immunol 7: 444–450.
-
7.
Jang MS, Sahastrabuddhe S, Yun C, Han SH, Yang JS, 2016. Serum bactericidal assay for the evaluation of typhoid vaccine using a semi-automated colony-counting method. Microb Pathog 97: 19–26.
-
8.
Liu X, Wang S, Sendi L, Caulfield MJ, 2004. High-throughput imaging of bacterial colonies grown on filter plates with application to serum bactericidal assays. J Immunol Methods 292: 187–193.
-
9.
Nahm MH, Yu J, Weerts HP, Wenzel H, Tamilselvi SC, Chandrasekaran L, Pasetti MF, Mani S, Kaminski RW, 2018. Development, Interlaboratory evaluations, and application of a simple, high-throughput Shigella serum bactericidal assay. mSphere 3: 1–14.
-
10.
Weerts HP, Yu J, Kaminski RW, Nahm MHA, 2019. A high-throughput Shigella-specific bactericidal assay. J Vis Exp 144: 1–9.
-
11.
Yang JS, Kim HJ, Yun C, Kang S, Im J, Kim H, Han SH, 2007. A semi-automated vibriocidal assay for improved measurement of cholera vaccine-induced immune responses. J Microbiol Methods 71: 141–146.
-
12.
Ayalew S, Confer AW, Shrestha B, Payton ME, 2012. A rapid microtiter plate serum bactericidal assay method for determining serum complement-mediated killing of Mannheimia haemolytica. J Microbiol Methods 89: 99–101.
-
13.
Sharma SK, Fatma T, Thukral SS, 1999. A simple and rapid serum bactericidal assay and its evaluation in clinical isolates of Klebsiella pneumoniae. J Microbiol Methods 39: 45–48.
-
14.
Rouphael NG, Satola S, Farley MM, Rudolph K, Schmidt DS, Gomez-de-leo P, Robbins JB, Schneerson R, Carlone GM, Romero-Steiner S, 2011 . Evaluation of serum bactericidal antibody assays for Haemophilus influenzae serotype a. Clin Vaccine Immunol 18: 243–247.
-
15.
Mountzouros KT, Howell AP, 2000. Detection of complement-mediated antibody-dependent bactericidal activity in a fluorescence-based serum bactericidal assay for group B Neisseria meningitidis. J Clin Microbiol 38: 2878–2884.
-
16.
Romero-Steiner S, Spear W, Brown N, Holder P, Hennessy T, de Leon PG, Carlone GM, 2004. Measurement of serum bactericidal activity specific for Haemophilus influenzae type b by using a chromogenic and fluorescent metabolic indicator. Clin Diagn Lab Immunol 11: 89–93.
-
17.
Rodriguez T, Lastre M, Cedre B, del Campo J, Bracho G, Zayas C, Taboada C, Diaz M, Sierra G, Perez O, 2002. Standardization of Neisseria meningitidis serogroup B colorimetric serum bactericidal assay. Clin Diagn Lab Immunol 9: 109–114.
-
18.
Necchi F, Saul A, Rondini S, 2017. Development of a high-throughput method to evaluate serum bactericidal activity using bacterial ATP measurement as survival readout. PLoS One 12: e0172163.
-
19.
Necchi F, Saul A, Rondini S, 2018. Setup of luminescence-based serum bactericidal assay against Salmonella Paratyphi A. J Immunol Methods 461: 117–121.
-
20.
Rossi O, Molesti E, Saul A, Giannelli C, Micoli F, Necchi F, 2020. Intra-laboratory evaluation of luminescence based high-throughput serum bactericidal assay (L-SBA) to determine bactericidal activity of human sera against Shigella .High Throughput 9: 1–12.
-
21.
Clow F, O’Hanlon CJ, Christodoulides M, Radcliff FJ, 2019. Feasibility of using a luminescence-based method to determine serum bactericidal activity against Neisseria gonorrhoeae. Vaccines (Basel) 7: 191.
-
22.
Jacobs AC, Didone L, Jobson J, Sofia MK, Krysan D, Dunman M, 2013. Adenylate kinase release as a high-throughput-screening-compatible reporter of bacterial lysis for identification of antibacterial agents. Antimicrob Agents Chemother 57: 26–36.
-
23.
Dzeja P, Terzic A, 2009. Adenylate kinase and AMP signaling networks: metabolic monitoring, signal communication and body energy sensing. Int J Mol Sci 10: 1729–1772.
-
24.
Ali M, Nelson AR, Lopez AL, Sack DA, 2015. Updated global burden of cholera in endemic countries. PLoS Negl Trop Dis 9: e0003832.
-
25.
Attridge SR, Johansson C, Trach DD, Qadri F, 2002. Sensitive microplate assay for detection of bactericidal antibodies to Vibrio cholerae O139. Clin Diagn Lab Immunol 9: 383–387.
-
26.
Azman AS et al., 2019. Estimating cholera incidence with cross-sectional serology. Sci Transl Med 11: 1–11.
-
27.
Richie E et al., 2000. Efficacy trial of single-dose live oral cholera vaccine CVD 103-HgR in North Jakarta, Indonesia, a cholera-endemic area. Vaccine 18: 2399–2410.
-
28.
Tacket CO et al., 1999. Randomized, double-blind, placebo-controlled, multicentered trial of the efficacy of a single dose of live oral cholera vaccine CVD 103-HgR in preventing cholera following challenge with Vibrio cholerae O1 El Tor Inaba three months after vaccination. Infect Immun 67: 6341–6345.
-
29.
Boutonnier A, Dassy B, Dumenil R, Guenole A, Ratsitorahina M, Migliani R, Fournier J, 2003. A simple and convenient microtiter plate assay for the detection of bactericidal antibodies to Vibrio cholerae O1 and Vibrio cholerae O139. J Microbiol Methods 55: 745–753.
-
30.
Kauffman RC et al., 2016. Single-cell analysis of the plasmablast response to Vibrio cholerae demonstrates expansion of cross-reactive memory B cells. mBio 7: 1–11.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1147 | 555 | 37 |
| Full Text Views | 207 | 12 | 0 |
| PDF Downloads | 159 | 14 | 0 |