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第三代检测甲型肝炎病毒(HAV)抗体试验需要特异性强的抗原。鉴于HAV在体外培养较为困难,作者建立了一种二步蔗糖离心法从病人粪便中提取HAV抗原的技术。该HAV抗原用于酶免疫试验(EIA)测定HAV抗体(IgM和IgG),具有很好的免疫化学活性,可以稀释40～150倍后使用。第一步取病人的HAV阳性粪便,制成20%的抽提物,以2500转/分,20℃离心1小时。取沉渣再次抽提可以提高HAV产量。将上清通过40%蔗糖溶液离心(7500转/分,3小
The third generation of Hepatitis A virus (HAV) antibody tests require specific antigens. In view of the difficulty of culturing HAV in vitro, the authors established a two-step sucrose centrifugation technique to extract HAV antigens from patient’s feces. This HAV antigen is used in Enzyme Immunoassay (EIA) for the determination of HAV antibodies (IgM and IgG) with good immunochemical activity and can be diluted 40 to 150 times before use. The first step to take the patient’s HAV positive stool, made of 20% of the extract, at 2500 r / min, 20 ° C for 1 hour. Taking the sediment again can improve the HAV yield. The supernatant was centrifuged through 40% sucrose solution (7500 rpm, 3 min