Interlaboratory study to assess precision and reproducibility of the meningococcal antigen surface expression (MEASURE) assay to quantify factor H binding protein expression at the surface of meningococcal serogroup B strains

IF 2.1 4区医学 Q3 INFECTIOUS DISEASES

Diagnostic microbiology and infectious disease Pub Date : 2025-05-30 DOI:10.1016/j.diagmicrobio.2025.116920

Jakob Loschko , Paul Liberator , Jamie Findlow , Jason Yip , Charles Tan , Karen Garcia , MaryAnn Murillo , Yamini Gorantla , Kimberly M. Moss , Panagiotis Maniatis , Stephen A. Clark , Ray Borrow

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引用次数: 0

Abstract

Background

The serum bactericidal antibody using human complement (hSBA) assay, the accepted surrogate measure of meningococcal vaccine efficacy, is limited by human sera and complement requirements. Pfizer developed and validated the flow-cytometry−based Meningococcal Antigen Surface Expression (MEASURE) assay to quantify surface-expressed factor H binding protein (fHbp) levels on intact meningococci. Surface expression of fHbp is correlated with hSBA assay killing by MenB-fHbp (Trumenba^Ⓡ)–induced antibody, meaning the MEASURE assay can be used to predict meningococcal serogroup B (MenB) strain susceptibility to antibodies elicited by MenB-fHbp. This study aimed to evaluate interlaboratory precision and reproducibility of the MEASURE assay.

Methods

The MEASURE assay was transferred to UK Health Security Agency (UKHSA) and US Centers for Disease Control and Prevention (CDC) laboratories. MEASURE assay results from 42 MenB strains encoding sequence-diverse fHbp variants that express fHbp at different levels were compared between the UKHSA, CDC, and Pfizer laboratories. Intermediate precision within each laboratory was determined.

Results

Pairwise comparisons of fHbp expression levels for all 42 MenB test strains showed >97 % agreement across the 3 laboratories when strains were grouped above or below a mean fluorescence intensity level of 1000, the threshold previously established as indicative of susceptibility to MenB-fHbp–induced antibodies in the hSBA assay. Each laboratory met assay precision criteria of ≤30 % total relative standard deviation.

Conclusions

Quantification of fHbp surface expression using the MEASURE assay is robust and reproducible across different laboratories. Previously determined cutoffs corresponding to predicted susceptibility to vaccine-induced antibodies can be applied to MEASURE data generated across laboratories.

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实验室间研究评估脑膜炎球菌抗原表面表达（MEASURE）测定定量脑膜炎球菌血清B组菌株表面因子H结合蛋白表达的准确性和重复性

使用人补体（hSBA）测定血清杀菌抗体是公认的脑膜炎球菌疫苗效力的替代测量方法，但受人血清和补体需求的限制。辉瑞公司开发并验证了基于流式细胞术的脑膜炎球菌抗原表面表达（MEASURE）测定，以定量完整脑膜炎球菌表面表达因子H结合蛋白（fHbp）水平。fHbp的表面表达与MenB-fHbp （TrumenbaⓇ）诱导抗体在hSBA试验中的杀死相关，这意味着MEASURE试验可用于预测脑膜炎球菌血清B组（MenB -fHbp）菌株对MenB-fHbp诱导抗体的敏感性。本研究旨在评价MEASURE测定法的实验室间精密度和重复性。方法将MEASURE方法转移到英国卫生安全局（UKHSA）和美国疾病控制与预防中心（CDC）实验室。在UKHSA， CDC和Pfizer实验室比较了42株编码不同序列的fHbp变体的MenB菌株的MEASURE检测结果，这些菌株表达不同水平的fHbp。确定了每个实验室的中间精密度。结果对所有42株MenB试验菌株的fHbp表达水平进行的双组比较显示，当菌株被分组在高于或低于平均荧光强度水平1000时，3个实验室的fHbp表达水平的一致性为97%，平均荧光强度水平1000是先前在hSBA试验中确定的对MenB-fHbp诱导抗体敏感的指示值。各实验室均满足≤30%总相对标准偏差的测定精密度标准。结论采用MEASURE法定量测定fHbp表面表达在不同实验室具有稳健性和可重复性。先前确定的与预测的疫苗诱导抗体易感性相对应的截止值可应用于跨实验室生成的MEASURE数据。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Diagnostic microbiology and infectious disease 医学-传染病学

CiteScore

5.30

自引率

3.40%

发文量

149

审稿时长

56 days

期刊介绍： Diagnostic Microbiology and Infectious Disease keeps you informed of the latest developments in clinical microbiology and the diagnosis and treatment of infectious diseases. Packed with rigorously peer-reviewed articles and studies in bacteriology, immunology, immunoserology, infectious diseases, mycology, parasitology, and virology, the journal examines new procedures, unusual cases, controversial issues, and important new literature. Diagnostic Microbiology and Infectious Disease distinguished independent editorial board, consisting of experts from many medical specialties, ensures you extensive and authoritative coverage.