Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i>
Development of a vaccine to limit the impact of antibiotic resistant <i>Neisseria gonorrhoeae</i> is now a global priority. Serum bactericidal antibody (SBA) is a possible indicator of protective immunity to <i>N. gonorrhoeae</i>, but conventional assays measure colony formin...
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2019-11-01
|
| Series: | Vaccines |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2076-393X/7/4/191 |
| _version_ | 1798000004469620736 |
|---|---|
| author | Fiona Clow Conor J O’Hanlon Myron Christodoulides Fiona J Radcliff |
| author_facet | Fiona Clow Conor J O’Hanlon Myron Christodoulides Fiona J Radcliff |
| author_sort | Fiona Clow |
| collection | DOAJ |
| description | Development of a vaccine to limit the impact of antibiotic resistant <i>Neisseria gonorrhoeae</i> is now a global priority. Serum bactericidal antibody (SBA) is a possible indicator of protective immunity to <i>N. gonorrhoeae</i>, but conventional assays measure colony forming units (CFU), which is time-consuming. A luminescent assay that quantifies ATP as a surrogate measure of bacterial viability was tested on <i>N. gonorrhoeae</i> strains FA1090, MS11 and P9-17 and compared to CFU-based readouts. There was a linear relationship between CFU and ATP levels for all three strains (<i>r </i>> 0.9). Normal human serum (NHS) is a common source of complement for SBA assays, but needs to be screened for non-specific bactericidal activity. NHS from 10 individuals were used for serum sensitivity assays—sensitivity values were significantly reduced with the ATP method for FA1090 (5/10, <i>p</i> < 0.05) and MS11 (10/10, <i>p</i> < 0.05), whereas P9-17 data were comparable for all donors. Our results suggest that measuring ATP underestimates serum sensitivity of <i>N. gonorrhoeae</i> and that the CFU method is a better approach. However, mouse anti-P9-17 outer membrane vesicles (OMV) SBA titres to P9-17 were comparable with both methods (<i>r </i>= 0.97), suggesting this assay can be used to rapidly screen sera for bactericidal antibodies to gonococci. |
| first_indexed | 2024-04-11T11:13:22Z |
| format | Article |
| id | doaj.art-e0a96a46339c44059c7b38c1baadf702 |
| institution | Directory Open Access Journal |
| issn | 2076-393X |
| language | English |
| last_indexed | 2024-04-11T11:13:22Z |
| publishDate | 2019-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Vaccines |
| spelling | doaj.art-e0a96a46339c44059c7b38c1baadf7022022-12-22T04:27:19ZengMDPI AGVaccines2076-393X2019-11-017419110.3390/vaccines7040191vaccines7040191Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i>Fiona Clow0Conor J O’Hanlon1Myron Christodoulides2Fiona J Radcliff3Department of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, University of Auckland, Auckland 1023, New ZealandDepartment of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, University of Auckland, Auckland 1023, New ZealandFaculty of Medicine, Academic Unit of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton, Southampton SO166YD, UKDepartment of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, University of Auckland, Auckland 1023, New ZealandDevelopment of a vaccine to limit the impact of antibiotic resistant <i>Neisseria gonorrhoeae</i> is now a global priority. Serum bactericidal antibody (SBA) is a possible indicator of protective immunity to <i>N. gonorrhoeae</i>, but conventional assays measure colony forming units (CFU), which is time-consuming. A luminescent assay that quantifies ATP as a surrogate measure of bacterial viability was tested on <i>N. gonorrhoeae</i> strains FA1090, MS11 and P9-17 and compared to CFU-based readouts. There was a linear relationship between CFU and ATP levels for all three strains (<i>r </i>> 0.9). Normal human serum (NHS) is a common source of complement for SBA assays, but needs to be screened for non-specific bactericidal activity. NHS from 10 individuals were used for serum sensitivity assays—sensitivity values were significantly reduced with the ATP method for FA1090 (5/10, <i>p</i> < 0.05) and MS11 (10/10, <i>p</i> < 0.05), whereas P9-17 data were comparable for all donors. Our results suggest that measuring ATP underestimates serum sensitivity of <i>N. gonorrhoeae</i> and that the CFU method is a better approach. However, mouse anti-P9-17 outer membrane vesicles (OMV) SBA titres to P9-17 were comparable with both methods (<i>r </i>= 0.97), suggesting this assay can be used to rapidly screen sera for bactericidal antibodies to gonococci.https://www.mdpi.com/2076-393X/7/4/191<i>neisseria gonorrhoeae</i>serum bactericidal activityluminescentatpserum-sensitivity |
| spellingShingle | Fiona Clow Conor J O’Hanlon Myron Christodoulides Fiona J Radcliff Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i> Vaccines <i>neisseria gonorrhoeae</i> serum bactericidal activity luminescent atp serum-sensitivity |
| title | Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i> |
| title_full | Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i> |
| title_fullStr | Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i> |
| title_full_unstemmed | Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i> |
| title_short | Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against <i>Neisseria gonorrhoeae</i> |
| title_sort | feasibility of using a luminescence based method to determine serum bactericidal activity against i neisseria gonorrhoeae i |
| topic | <i>neisseria gonorrhoeae</i> serum bactericidal activity luminescent atp serum-sensitivity |
| url | https://www.mdpi.com/2076-393X/7/4/191 |
| work_keys_str_mv | AT fionaclow feasibilityofusingaluminescencebasedmethodtodetermineserumbactericidalactivityagainstineisseriagonorrhoeaei AT conorjohanlon feasibilityofusingaluminescencebasedmethodtodetermineserumbactericidalactivityagainstineisseriagonorrhoeaei AT myronchristodoulides feasibilityofusingaluminescencebasedmethodtodetermineserumbactericidalactivityagainstineisseriagonorrhoeaei AT fionajradcliff feasibilityofusingaluminescencebasedmethodtodetermineserumbactericidalactivityagainstineisseriagonorrhoeaei |