Zanetti A R, Bedarida G, Ferroni P, D'Agostino F, Bianchi V
J Virol Methods. 1980 Dec;2(1-2):71-83. doi: 10.1016/0166-0934(80)90041-5.
Following Blumberg's discovery of hepatitis B surface antigen (HBsAg), many attempts have been made to develop several in vitro diagnostic techniques for the detection of this antigen and its homologous antibody. The two-dimensional micro-Ouchterlony immunodiffusion has been the first technique used, rapidly replaced by procedures of increasing sensitivity characterized as second-generation and the currently available third-phase tests which include radioimmunoassay (RIA), reverse passive haemagglutination (RPHA), reverse passive latex agglutination (RPLA) and enzyme immunoassay (EIA). Among these, RIA appears to be the most sensitive and specific, whereas EIA, RPHA and RPLA have the advantage of long shelf-life of stable reagents, no need for sophisticated and expensive equipment and no hazard associated with the handling of radioactive isotopes. Moreover, the sensitivity of EIA should increase by objective reading with a colorimeter. The most sensitive method for the detection of surface antibody (anti-HBs) is again RIA, whereas passive haemagglutination (PHA) had the advantage of providing titres. Finally EIA, based on inhibition of a known amount of HBsAg, has at least the same sensitivity as PHA, but has the advantage that reagents are more stable and that it permits screening for both HBsAg and anti-HBs with the same reagents at the same time. The application of these highly sensitive techniques for the detection of HBsAg and anti-HBs has resulted in a consistent reduction in the incidence of post-transfusion hepatitis type B and in a better understanding of the aetiology, epidemiology and natural history of this infection.
在布隆伯格发现乙型肝炎表面抗原（HBsAg）之后，人们进行了许多尝试，以开发多种体外诊断技术来检测这种抗原及其同源抗体。二维微量双向免疫扩散是最早使用的技术，但很快就被灵敏度更高的第二代检测方法以及目前可用的第三代检测方法所取代，第三代检测方法包括放射免疫分析（RIA）、反向被动血凝试验（RPHA）、反向被动乳胶凝集试验（RPLA）和酶免疫分析（EIA）。其中，RIA似乎是最灵敏和特异的，而EIA、RPHA和RPLA具有试剂稳定性好、保质期长、无需复杂昂贵设备以及不存在放射性同位素处理风险等优点。此外，通过比色计进行客观读数可提高EIA的灵敏度。检测表面抗体（抗-HBs）最灵敏的方法同样是RIA，而被动血凝试验（PHA）具有能够提供滴度的优势。最后，基于抑制已知量HBsAg的EIA，其灵敏度至少与PHA相同，但具有试剂更稳定的优点，并且可以使用相同试剂同时筛查HBsAg和抗-HBs。这些高灵敏度技术在检测HBsAg和抗-HBs方面的应用，使得输血后乙型肝炎的发病率持续下降，并且对这种感染的病因、流行病学和自然史有了更深入的了解。
