人免疫缺损病毒蛋白印迹法的改进

IMPROVEMENT OF WESTERN BLOT TEST FOR DETECTION OF ANTIBODY TO HUMAN IMMUNODEFICIENCY VIRUS ( HIV )

对传统的人免疫缺损病毒(HIV 1)蛋白印迹法(Western blot)进行了一系列改进。实验表明,用裂解的HIV感染细胞直接做蛋白印迹是完全可行的,所有HIV的主要蛋白带均清晰可见,无明显的非特异性着色.用吐温PBS液加小牛血清做封闭可以替代成份复杂的Cohen液。血清和酶标物的孵育时间可缩短到60～30分钟,使抗体检测可在2小时内完成.经过改进的蛋白印迹法在分离我国首株HIV和对HIV感染进行血清学诊断中,被证明是一种简便、高效、准确和可靠的实验方法,并在除HIV 1以外的其它病毒系统中,也显示了潜在的应用价值.

To overcome the disadvantages of the tranditional Western blot ( WB ) , such as complicated procedures,sophisticaed instrument and expensive reagents used in HIV antigen purification and aptibody detection, a series of improvements were made. The lysate of HlV infected cells was directly electrophoresed and transferred to nitrocellulose paper. The complicated Cohen buffer was substituted by PBS buffer containing calf serum and the total incubation time in antibody detection can be reduced to one hour. The results showed that all major HIV proteinbends can be clearly seen with a very low nonspecific background. It was proved that the improved WB is a hig hly effective, accurate and simple method which can be applied to the antigen identification and antibody detection for HIV,and possibly for other viruses.