Takahashi H, Kameda H, Wands J R
Gastrointestinal Unit, Massachusetts General Hospital, Boston 02114.
J Immunol Methods. 1988 Sep 13;112(2):191-200. doi: 10.1016/0022-1759(88)90357-2.
We have developed and employed a second generation monoclonal immunoradiometric assay (M2-IRMA) using antibodies of high affinity for epitopes that reside on hepatitis B surface antigen (HBsAg). This assay is capable of detecting as little as 15 pg/ml of HBsAg in serum. Improvements in sensitivity over a first generation immunoradiometric assay (MI-IRMA) was achieved by increasing the sample volume and time of incubation, and subjecting the reaction to a mechanical rotary devise. We then studied 164 subjects with chronic hepatitis, 105 with cirrhosis, 67 with hepatocellular carcinoma, six with acute hepatitis A, seven with acute hepatitis B, 167 chronic carriers of hepatitis B virus (HBV) and 235 healthy individuals from Japan and compared the results of the M2-IRMA to a conventional polyclonal radioimmunoassay (P-RIA). By using a more sensitive assay design (M2-IRMA), a significant number of additional cases of HBV infection heretofore unsuspected in the etiology of chronic liver disease were identified. We conclude that improvement in assay sensitivity for HBsAg is important in the serologic diagnosis of HBV in patients with chronic hepatitis, cirrhosis and hepatocellular carcinoma.
我们研发并应用了第二代单克隆免疫放射测定法（M2-IRMA），该方法使用了对乙型肝炎表面抗原（HBsAg）上的表位具有高亲和力的抗体。此测定法能够检测出血清中低至15 pg/ml的HBsAg。通过增加样本量和孵育时间，并使反应通过机械旋转装置进行，相对于第一代免疫放射测定法（MI-IRMA），其灵敏度得到了提高。然后，我们研究了来自日本的164例慢性肝炎患者、105例肝硬化患者、67例肝细胞癌患者、6例甲型肝炎患者、7例乙型肝炎患者、167例慢性乙型肝炎病毒（HBV）携带者以及235名健康个体，并将M2-IRMA的结果与传统的多克隆放射免疫测定法（P-RIA）进行了比较。通过使用更灵敏的测定设计（M2-IRMA），在慢性肝病病因中发现了大量此前未被怀疑的HBV感染新病例。我们得出结论，提高HBsAg测定的灵敏度对于慢性肝炎、肝硬化和肝细胞癌患者的HBV血清学诊断很重要。
