Rodrigo M J, Codina R, de Gracia J, Morell F, Pascual C
Servicio de Bioquímica (Unidad de Inmunología), Hospital General Universitario Vall d'Hebron, Universitat Autónoma de Barcelona.
Med Clin (Barc). 1992 Feb 8;98(5):166-70.
Quantification of the IgG subclasses (IgGS) requires highly sensitive and specific techniques since their molecular structure is more than 95% homologous. At present, ELISA and RIA are the most appropriate techniques; although the variability of these techniques and the possible ethnic differences in serum levels of IgGS oblige each laboratory to establish its own reference values in a normal population, a condition necessary for defining IgGS deficits.
In the present study the normal serum values of IgGS in 100 healthy individuals were established. Serum quantification of the IgGS was performed by optimization of the indirect ELISA technique using AcMo and OMS reference standards H00-03 for the 4 subclasses, calibrated vs the OMS WHO 67/97 pattern.
Normal values observed for IgG1 were 2.61-10.81 g/l; IgG2, 1.12-4.08 g/l; IgG3, 0.22-2.88 g/l; and IgG4, 0.05-1.56 g/l obtained as of the mean +/- 2 SD for IgG1 and as of the extent of the values of the remaining 3 subclasses by following a non-normal distribution. The sensitivity of the technique was 0.05 g/l with mean intra and inter analysis coefficients of 3.5% and 10.7% respectively. The correlation between total IgG and the sum of the 4 subclasses showed a r = 0.894 (p less than 0.01).
The ELISA technique using AcMo and OMS reference standards is an adequate method for the quantification of the IgG subclasses. However, given the variability of this technique and in order to avoid error, strict working conditions, such as the use of control sera at 2 concentration levels, should be observed for the 4 IgGS in each of the analytic series and each series should also be duplicated.
由于IgG亚类(IgGS)的分子结构同源性超过95%,因此对其进行定量需要高度敏感和特异的技术。目前,ELISA和RIA是最适用的技术;尽管这些技术存在变异性以及IgGS血清水平可能存在种族差异,这使得每个实验室都必须在正常人群中建立自己的参考值,这是定义IgGS缺乏的必要条件。
在本研究中,确定了100名健康个体的IgGS正常血清值。采用间接ELISA技术,使用AcMo和OMS参考标准品H00 - 03对4个亚类进行血清定量,以OMS WHO 67/97模式进行校准。
观察到IgG1的正常值为2.61 - 10.81 g/l;IgG2为1.12 - 4.08 g/l;IgG3为0.22 - 2.88 g/l;IgG4为0.05 - 1.56 g/l,IgG1取均值±2标准差,其余3个亚类取数值范围,因其呈非正态分布。该技术的灵敏度为0.05 g/l,平均批内和批间分析系数分别为3.5%和10.7%。总IgG与4个亚类之和的相关性显示r = 0.894(p < 0.01)。
使用AcMo和OMS参考标准品的ELISA技术是定量IgG亚类的合适方法。然而,鉴于该技术的变异性,为避免误差,在每个分析系列中对4种IgGS应严格遵守工作条件,如使用2种浓度水平的对照血清,并且每个系列都应重复。