Thomas H I, Morgan-Capner P
Department of Virology, Preston Infirmary.
Epidemiol Infect. 1988 Jun;100(3):443-54. doi: 10.1017/s0950268800067182.
A solid-phase antigen enzyme-linked immunosorbent assay (ELISA) was developed for the detection of rubella-specific IgG subclasses. For rubella-specific IgG1 and IgG3 sera were quantitated in arbitrary units (au) by comparison with standard curves. A concentration of 3 au was taken as that indicating positivity for specific IgG1 and specific IgG3. No sera reactive for specific IgG2 and IgG4 have been found, and thus the assay reagents were controlled by testing dilutions of a standard calibrant serum containing known concentrations of the specific IgG subclasses. Of 105 unselected sera negative for rubella antibody by radial haemolysis (RH), two gave concentrations of specific IgG1 greater than 3 au and both were positive by rubella latex agglutination (LA). The sensitivity of the assay for specific IgG1 was confirmed by examining 25 selected sera negative by RH but reactive by LA. Twenty-one gave concentrations greater than 3 au. None of these 130 was positive for specific IgG3. All 63 sera containing greater than 15 international units rubella antibody by RH from cases of rubella in the remote past contained specific IgG1 and eight contained specific IgG3. In 79 cases of primary rubella, specific IgG1 developed in all cases by day 8. Specific IgG3 became detectable in all cases except one by day 16. Serum taken on day 21 from one case was negative for specific IgG3 but the absence of later sera precluded further investigation. One case had become negative for specific IgG3 by day 56. Sera from 24 cases of rubella reinfection were examined and all contained specific IgG1. In three cases of symptomatic reinfection, specific IgG3 was detectable in two but not in the remaining case. In 2 of the 21 cases of asymptomatic reinfection only a very early or a very late serum was available. Of the remaining 19 cases, 7 had detectable specific IgG3. However, only one of 9 sera collected 30-50 days after contact contained specific IgG3. Thus for the asymptomatic patient for whom other serological tests suggest a recent rubella infection, the failure to detect specific IgG3 in sequential sera collected after contact suggests reinfection rather than primary rubella. The detection of specific IgG3 did not correlate with the presence of specific IgM. Sera collected 6-8 weeks after rubella vaccination had detectable specific IgG1 in 32 of 33 cases and specific IgG3 in 9 of 33. The remaining vaccinee was seronegative.
开发了一种用于检测风疹特异性IgG亚类的固相抗原酶联免疫吸附测定(ELISA)。对于风疹特异性IgG1和IgG3,通过与标准曲线比较以任意单位(au)对血清进行定量。将3 au的浓度视为表明特异性IgG1和特异性IgG3呈阳性的浓度。未发现对特异性IgG2和IgG4有反应的血清,因此通过检测含有已知浓度特异性IgG亚类的标准校准血清的稀释液来控制测定试剂。在105份通过放射溶血(RH)检测风疹抗体为阴性的未选择血清中,两份血清的特异性IgG1浓度大于3 au,且两份血清通过风疹乳胶凝集试验(LA)均为阳性。通过检查25份经RH检测为阴性但经LA检测有反应的选择血清,证实了该测定对特异性IgG1的敏感性。21份血清的浓度大于3 au。这130份血清中没有一份特异性IgG3呈阳性。所有63份来自既往风疹病例且通过RH检测风疹抗体大于15国际单位的血清均含有特异性IgG1,8份含有特异性IgG3。在79例原发性风疹病例中,所有病例在第8天均出现特异性IgG1。除1例病例外,所有病例在第16天均可检测到特异性IgG3。从1例病例在第21天采集的血清中特异性IgG3为阴性,但由于没有后续血清,无法进行进一步调查。1例病例在第56天时特异性IgG3变为阴性。检查了24例风疹再感染病例的血清,所有血清均含有特异性IgG1。在3例有症状的再感染病例中,2例可检测到特异性IgG3,另1例未检测到。在21例无症状再感染病例中,有2例仅获得了非常早期或非常晚期的血清。在其余19例病例中,7例可检测到特异性IgG3。然而,在接触后30 - 50天采集的9份血清中,只有1份含有特异性IgG3。因此,对于其他血清学检测提示近期风疹感染的无症状患者,接触后连续采集的血清中未检测到特异性IgG3提示为再感染而非原发性风疹。特异性IgG3的检测与特异性IgM的存在无关。在风疹疫苗接种后6 - 8周采集的血清中,33例中有32例可检测到特异性IgG1,33例中有9例可检测到特异性IgG3。其余接种者血清学阴性。