Ireland D, Samuel D
Division of Microbiological Reagents and Quality Control, Central Public Health Laboratory, Colindale, London, UK.
J Biolumin Chemilumin. 1989 Jul;4(1):159-63. doi: 10.1002/bio.1170040124.
A chemiluminescent enzyme linked immunosorbent assay (ELISA) for the detection of antibody to hepatitis B virus surface antigen (anti-HBs) in human serum has been developed. Polystyrene microtitre plates were coated with recombinant, yeast-derived hepatitis B surface antigen (rec-HBsAg). Patient serum samples and appropriate controls were added to the rec-HBsAg-coated wells and incubated to bind anti-HBs. The wells were then washed and a fluorescein isothiocyanate (FITC) conjugate of a human plasma-derived hepatitis B surface antigen (HBsAg) was added. Following incubation and further washing, the bound FITC-labelled HBsAg was detected after addition of a horseradish peroxidase (HRP) conjugate of a monoclonal anti-FITC antibody and assaying for the enzyme. The activity of the HRP was measured using luminol and hydrogen peroxide as substrates and iodophenol as a chemiluminescence enhancer. The luminescence was recorded using a camera luminometer. Preliminary tests have shown the assay to be suitable for the detection of antibody in sera from both vaccinees and also from individuals with a past hepatitis B virus infection. The use of the FITC-anti-FITC system together with the measurement of a chemiluminescence signal makes possible the completion of this assay in a few hours. The assay has been shown to be both specific and sensitive and provides a permanent photographic record.
已开发出一种化学发光酶联免疫吸附测定法（ELISA），用于检测人血清中乙型肝炎病毒表面抗原抗体（抗-HBs）。聚苯乙烯微量滴定板用重组酵母源乙型肝炎表面抗原（rec-HBsAg）包被。将患者血清样本和适当的对照加入到包被有rec-HBsAg的孔中并孵育，以使抗-HBs结合。然后洗涤孔，加入人血浆源乙型肝炎表面抗原（HBsAg）的异硫氰酸荧光素（FITC）缀合物。孵育并进一步洗涤后，加入单克隆抗FITC抗体的辣根过氧化物酶（HRP）缀合物并测定该酶后，检测结合的FITC标记的HBsAg。使用鲁米诺和过氧化氢作为底物以及碘苯酚作为化学发光增强剂来测量HRP的活性。使用相机光度计记录发光情况。初步测试表明该测定法适用于检测来自疫苗接种者以及既往感染过乙型肝炎病毒个体的血清中的抗体。FITC-抗FITC系统的使用以及化学发光信号的测量使得该测定法能够在数小时内完成。该测定法已被证明具有特异性和敏感性，并提供永久的照片记录。
