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使用基于发光的方法测定血清对……的杀菌活性的可行性

Feasibility of Using a Luminescence-Based Method to Determine Serum Bactericidal Activity against .

作者信息

Clow Fiona, O'Hanlon Conor J, Christodoulides Myron, Radcliff Fiona J

机构信息

Department of Molecular Medicine and Pathology, Faculty of Medical and Health Sciences, University of Auckland, Auckland 1023, New Zealand.

Faculty of Medicine, Academic Unit of Clinical and Experimental Sciences, Sir Henry Wellcome Laboratories, University of Southampton, Southampton SO166YD, UK.

出版信息

Vaccines (Basel). 2019 Nov 21;7(4):191. doi: 10.3390/vaccines7040191.

Abstract

Development of a vaccine to limit the impact of antibiotic resistant is now a global priority. Serum bactericidal antibody (SBA) is a possible indicator of protective immunity to , 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 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 ( > 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, < 0.05) and MS11 (10/10, < 0.05), whereas P9-17 data were comparable for all donors. Our results suggest that measuring ATP underestimates serum sensitivity of 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 ( = 0.97), suggesting this assay can be used to rapidly screen sera for bactericidal antibodies to gonococci.

摘要

研发一种限制抗生素耐药性影响的疫苗现已成为全球优先事项。血清杀菌抗体(SBA)可能是对淋病奈瑟菌保护性免疫的一个指标,但传统检测方法测量的是菌落形成单位(CFU),这很耗时。一种将ATP定量作为细菌活力替代指标的发光检测方法在淋病奈瑟菌菌株FA1090、MS11和P9 - 17上进行了测试,并与基于CFU的读数进行了比较。所有三种菌株的CFU与ATP水平之间均呈线性关系(>0.9)。正常人血清(NHS)是SBA检测中补体的常见来源,但需要筛查其非特异性杀菌活性。来自10名个体的NHS用于血清敏感性检测——对于FA1090(5/10,<0.05)和MS11(10/10,<0.05),ATP方法的敏感性值显著降低,而P9 - 17的数据在所有供体中具有可比性。我们的结果表明,测量ATP会低估淋病奈瑟菌的血清敏感性,CFU方法是更好的方法。然而,小鼠抗P9 - 17外膜囊泡(OMV)对P9 - 17的SBA效价在两种方法中具有可比性(=0.97),这表明该检测方法可用于快速筛查血清中针对淋病奈瑟菌的杀菌抗体。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7f6b/6963289/893cc36c8c08/vaccines-07-00191-g001.jpg

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