Densitometric analysis of Western blot assays for feline immunodeficiency virus antibodies
Densitometric analysis of Western blot assays for feline immunodeficiency virus antibodies
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DOI：
10.1016/s0165-2427(01)00265-3
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发表时间：
2001-05-30
影响因子：
1.8
通讯作者：
Poli, A
中科院分区：
文献类型：
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作者：
Calandrella, M;Matteucci, D;Poli, A
用半自动光密度计反射光密度法分析了猫免疫缺陷病毒（FIV）抗体的蛋白质印迹（WB）条带。该方法用于定量接种疫苗的猫和自然或实验感染的猫中对不同FIV蛋白的抗体应答。为了提高重现性，对试剂和方案进行了准确标准化，并添加了内部对照。在第一种形式中，将由猫IgG组成的内部对照条带添加到每个印迹中，以使条带强度变化的影响最小化。在第二种形式中，根据测试血清和阳性和阴性标准血清产生的密度的比率计算抗体浓度。用标准的酶联免疫吸附试验（ELISA）检测血清，并与相应的WB结果进行比较。两种方法所得结果之间存在统计学显著正相关，当ELISA滴度与校正WB值比较时，这一点尤其明显（P = 0.001）。WB测定的光密度分析允许量化针对FIV蛋白的抗体，并且可能有助于调查在FIV疫苗接种研究中保护的可能的体液免疫相关性和在感染的早期阶段的抗体产生。Gag和Env FIV抗原抗体的定量可用于获得关于FIV疾病过程的进一步信息，如先前在人类免疫缺陷病毒-1（HIV-1）感染中所证明的。(C)2001 Elsevier Science B. V.保留所有权利。
Western blot (WB) strips for antibodies directed to feline immunodeficiency virus (FIV) were analysed using reflectance densitometry by a semiautomatic densitometer. This method was used to quantify the antibody responses to different FIV proteins in both vaccinated and naturally or experimentally-infected cats. In order to increase reproducibility, reagents and protocols were accurately standardised and internal controls were added. In a first format, an internal control band consisting of feline IgG was added to each blot to minimise the effect of band intensity variation. In a second format, antibody concentrations were calculated from the ratio of the dens ities produced by test sera and by positive and negative standard sera. The sera under scrutiny were also examined by standard enzyme-linked immunosorbent assay (ELISA) and the results obtained compared with those of the corresponding WB. A statistically significant positive correlation was found between the results obtained with the two methods, and this was especially evident when ELISA titres were compared to corrected WB values (P = 0.001). Densitometric analysis of WB assays allowed to quantify the antibodies against FIV proteins and might be useful to investigate possible humoral immune correlates of protection in FIV vaccination studies and antibody production in the early phase of infection. The quantitation of antibodies to Gag and Env FIV antigens might be used to obtain further informations on the course of FIV disease, as previously demonstrated in human immunodeficiency virus-1 (HIV-1) infections. (C) 2001 Elsevier Science B.V. All rights reserved.