Banfi N H, Minervini M V, Scoccia A E
Servicio de Laboratorio Central, Hospital I.A.C. San Juan de Dios, La Plata, Argentina.
Rev Argent Microbiol. 1995 Jul-Sep;27(3):123-9.
The aim of this work was to increase sensitivity in the detection of antigens from HIV-infected patients, through a process of immune complex dissociation without loss of antigenicity. 500 microliters of sera were mixed with 100 microliters of PEG 12%, stored one night in refrigerator, and centrifuged at 2000 g during 20 minutes. 200 microliters of buffer AcH/Ac- (pH 3.5) were added to the sediment, and incubated at 37 degrees C during one hour with periodic shaking. This was neutralized with 100 microliters of buffer TRIS/CIH (pH 8.6). The antigen was investigated in the original sample, supernatant and sediment. Samples of 105 patients with positive serology, confirmed by Western Blot following CDC criteria, were processed. The antigen was detected in 62 (59%) samples precipitated with PEG, but only 35 (33%) when conventional methods were used. Applying statistics X2: 13.97, P < 0.001, a highly significant association can be observed between PEG dissociation treatment and antigen detection. 27 negative sera by the standard method became positive in the whole sediment, and only 8 in the supernatant. In addition, 40 negative sera were processed, which had not become positive for the antigen by PEG treatment.
这项工作的目的是通过免疫复合物解离过程且不损失抗原性,提高对HIV感染患者抗原检测的灵敏度。将500微升血清与100微升12%的聚乙二醇(PEG)混合,在冰箱中储存一晚,然后以2000克离心20分钟。向沉淀物中加入200微升乙酸/乙酸盐缓冲液(pH 3.5),在37摄氏度下孵育1小时并定期振荡。用100微升三羟甲基氨基甲烷/氯化氢缓冲液(pH 8.6)将其中和。对原始样本、上清液和沉淀物中的抗原进行检测。对105例血清学阳性患者的样本按照疾病控制与预防中心(CDC)标准经蛋白质印迹法确认后进行处理。用PEG沉淀的样本中有62例(59%)检测到抗原,而采用传统方法时仅35例(33%)检测到。应用统计学方法卡方检验:X2为13.97,P < 0.001,可以观察到PEG解离处理与抗原检测之间存在高度显著的关联。27例标准方法检测为阴性的血清在整个沉淀物中变为阳性,而上清液中仅有8例。此外,对40例经PEG处理后抗原仍未呈阳性的阴性血清进行了处理。