Maslanka S E, Gheesling L L, Libutti D E, Donaldson K B, Harakeh H S, Dykes J K, Arhin F F, Devi S J, Frasch C E, Huang J C, Kriz-Kuzemenska P, Lemmon R D, Lorange M, Peeters C C, Quataert S, Tai J Y, Carlone G M
Childhood and Respiratory Diseases Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
Clin Diagn Lab Immunol. 1997 Mar;4(2):156-67. doi: 10.1128/cdli.4.2.156-167.1997.
A standardized serum bactericidal assay (SBA) is required to evaluate the functional activity of antibody produced in response to Neisseria meningitidis serogroup A and C vaccines. We evaluated assay parameters (assay buffer, target strains, growth of target cells, target cell number, complement source and concentration, and methods for growth of surviving bacteria) which may affect the reproducibility of SBA titers. The various assay parameters and specificity of anticapsular antibody to five serogroup A strains (A1, ATCC 13077, F8238, F9205, and F7485) and four serogroup C strains (C11, G7880, G8050, and 1002-90) were evaluated with Centers for Disease Control and Prevention meningococcal quality control sera. The critical assay parameters for the reproducible measurement of SBA titers were found to include the target strain, assay incubation time, and complement. The resulting standardized SBA was used by 10 laboratories to measure functional anticapsular antibody against serogroup A strains F8238 and serogroup C strain C11. In the multilaboratory study, SBA titers were measured in duplicate for 14 pairs of sera (seven adults and seven children) before and after immunization with a quadrivalent polysaccharide (A, C, Y, and W-135) vaccine. The standardized SBA was reliable in all laboratories regardless of experience in performing SBAs. For most sera, intralaboratory reproducibility was +/- 1 dilution; interlaboratory reproducibility was +/- 2 dilutions. The correlation between median titers (interlaboratory) and enzyme-linked immunosorbent assay total antibody concentrations was high for both serogroup A (r = 0.86; P < 0.001; slope = 0.5) and serogroup C (n = 0.86; P < 0.001; slope = 0.7). The specified assay, which includes the critical parameters of target strain, incubation time, and complement source, will facilitate interlaboratory comparisons of the functional antibody produced in response to current or developing serogroup A and C meningococcal vaccines.
需要一种标准化血清杀菌试验（SBA）来评估针对A群和C群脑膜炎奈瑟菌疫苗产生的抗体的功能活性。我们评估了可能影响SBA效价重复性的试验参数（试验缓冲液、靶菌株、靶细胞生长、靶细胞数量、补体来源和浓度以及存活细菌生长方法）。使用美国疾病控制与预防中心的脑膜炎球菌质量控制血清评估了各种试验参数以及抗荚膜抗体对5株A群菌株（A1、ATCC 13077、F8238、F9205和F7485）和4株C群菌株（C11、G7880、G8050和1002 - 90）的特异性。发现可重复测量SBA效价的关键试验参数包括靶菌株、试验孵育时间和补体。10个实验室使用所得的标准化SBA来测量针对A群菌株F8238和C群菌株C11的功能性抗荚膜抗体。在多实验室研究中，在使用四价多糖（A、C、Y和W - 135）疫苗免疫前后，对14对血清（7名成人和7名儿童）的SBA效价进行了一式两份测量。无论各实验室进行SBA的经验如何，标准化SBA在所有实验室中都是可靠的。对于大多数血清，实验室内重复性为±1个稀释度；实验室间重复性为±2个稀释度。A群（r = 0.86；P < 0.001；斜率 = 0.5）和C群（r = 0.86；P < 0.001；斜率 = 0.7）的中位数效价（实验室间）与酶联免疫吸附测定总抗体浓度之间的相关性都很高。包括靶菌株、孵育时间和补体来源等关键参数的特定试验，将有助于实验室间比较针对当前或正在研发的A群和C群脑膜炎球菌疫苗产生的功能性抗体。
